Histone demethylase inhibitors

ABSTRACT

The present embodiments provide for substituted triazolylpyridine derivative compounds, and pharmaceutical compositions comprising said compounds. The subject compounds and compositions are useful for modulating the activity of histone demethylase enzymes. Additionally, the subject compounds and compositions are useful for the treatment of cancer or other neoplastic diseases, or maladies associated with abnormal histone demethylase activity. Accordingly, the substituted triazolylpyridine derivative compounds described herein are useful in methods and medicaments for treating cancer.

RELATED APPLICATIONS

This Application claims priority benefit of U.S. Application No.62/324,813 filed Apr. 19, 2016, and incorporated entirely herein for allpurposes.

FIELD

The present embodiments relate generally to compounds, pharmaceuticalcompositions, and methods for modulating the activity of histonedemethylases.

BACKGROUND

A need exists in the art for an effective treatment of cancer,neoplastic diseases, or other maladies associated with histonedemethylase activity.

SUMMARY

The present embodiments provide substituted triazolylpyridine derivativecompounds and pharmaceutical compositions comprising said compounds. Thesubject compounds and compositions are useful for the inhibition ofhistone demethylases. Furthermore, the subject compounds andcompositions are useful for the treatment of cancer, such as pancreaticcancer, prostate cancer, breast cancer, bladder cancer, lung cancer,gastric cancer, leukemia and/or melanoma and the like. The substitutedtriazolylpyridine derivatives are based upon a di-substituted pyridinering bearing at the 4-position a substituted triazolyl group which isthe acid bioisostere and at the 3-position a substituted amine group orat the 2-position a substituted 1-pyrazolyl group.

At least one embodiment provides a compound having the structure ofFormula I

-   -   wherein the compound of Formula I includes a pharmaceutically        acceptable salt thereof, and    -   wherein    -   R¹ is halogen, —CH₂G, —NHG, or —OG, in which G is —X—Y, wherein        X is hydrogen or C₁ alkyl, and Y is optionally substituted        aralkyl, aralkenyl, aralkynyl, carbocyclyl, carbocyclylalkyl,        heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl;    -   R² is halogen or CF₃; and    -   R⁵ is hydrogen, methyl, ethyl, isoproplyl, t-butyl, —CHF₂,        —CH₂F, —CF₃, —CH₂OH, —CHCH₃OH, or —C(CH₃)₂OH.

In some embodiments of the compound of Formula I, R¹ is fluorine.

In some embodiments of the compound of Formula I, R² is —CF₃.

In some embodiments of the compound of Formula I, R⁵ is hydrogen.

In some embodiments of Formula I, Y is optionally substituted adamantyl,benzofuranyl, 2,3-dihydrobenzofuranyl, chromanyl, indanyl, indolyl,naphthyl, 1,2-dihydronaphthyl, phenyl, pyridyl, tetrahydroquinolinyl,tetralinyl, 2,3-dihydrobenzo[b]-[1,4]dioxinyl, or thiochromanyl.

In some embodiments of the compound of Formula I, Y is phenylsubstituted with alkyl, alkynyl, chloro, fluoro, fluoroalkyl, nitro; oroptionally substituted aralkyl, aralkenyl, aralkynyl, carbocyclyl,carbocyclylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl orheteroarylalkyl; or —R^(b)—OR^(a), —R^(b)—OC(O)—R^(a),—R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))_(2, —R)^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a), —R^(b)—OvR^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a), —R^(b)—S(O)_(t)OR^(a),—R^(b)—S(O)_(t)OR^(a), or —R^(b)—S(O)_(t)N(R^(a))₂, in which each R^(a)is independently hydrogen, alkyl, fluoroalkyl, cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl, or heteroarylalkyl; each R^(b) is independently a directbond or a straight or branched alkyl chain; each R^(c) is a straight orbranched alkyl chain; and t is 1 or 2.

In at least one embodiment, the compound of Formula I has the structure:

-   -   wherein G, R², and R⁵ are as described above.

In some embodiments of a compound of Formula Ia, R² is CF₃.

In some embodiments of a compound of Formula Ia, R⁵ is hydrogen.

In some embodiments of a compound of Formula Ia, G is —X—Y, wherein X ishydrogen and Y is disubstituted phenyl, wherein the substituents arehalogens.

In some embodiments, the compound of Formula I has the structure:

-   -   wherein R² and R⁵ are as described above; and    -   Z is independently at least one hydrogen, halogen, —OH, —CN, or        optionally substituted C₁-C₆ alkyl, C₁-C₆ alkoxy, C₃-C₇        carbocyclyl, C₃-C₇ carbocyclyloxy, C₄-C₁₂ carbocyclylalkyl,        C₄-C₁₂ carbocyclylalkoxy, C₁-C₆ alkynyl, C₁-C₆ alkenyl, C₆-C₁₀        aryl, C₆-C₁₀ aryloxy, C₆-C₁₀ aryl-S—, C₇-C₁₄ aralkoxy,        heteroaryl, or heteroaryloxy.

In some embodiments of the compound of Formula Ib, Z is at least onehalogen.

In some embodiments of the compound of Formula Ib, Z is at least onefluoro.

In other embodiments of the compound of Formula Ib, Z is independentlyat least one hydrogen, halogen, cyano, NH₂, NHR^(d), N(R^(d))₂,NHC(O)R^(d), NHC(O)OR^(d), NHC(O)NHR^(d), NHC(O)N(R^(d))₂, NHS(O)₂R^(d),NR^(d)C(O)R^(d), NR^(d)C(O)OR^(d), NR^(d)C(O)NHR^(d),NR^(d)C(O)N(R^(d))₂, NR^(d)S(O)₂R^(d), in which each R^(d) isindependently alkyl, aryl, aralkyl, carbocyclyl, heterocyclyl,heteroaryl, carbocyclylalkyl, heterocyclylalkyl, or heteroarylalkyl; orZ is independently at least one optionally substituted alkyl, alkenyl,alkynyl, alkoxy, aryl, aryloxy, aralkyl, carbocyclyl, heterocyclyl,heteroaryl, carbocyclylalkyl, heterocyclylalkyl, or heteroarylalkyl.

In some embodiments, R² is CF₃.

In some embodiments, R⁵ is hydrogen.

In some embodiments, Z is two halogens, which may be at least one F orCl.

In some embodiments, Z is optionally substituted C₄-C₁₂carbocyclylalkyl.

At least one embodiment provides a compound having the structure ofFormula II

-   -   wherein a compound of Formula II includes a pharamceutical salt        thereof, and wherein    -   R² is halogen or CF₃;    -   R³ is hydrogen, halogen, —OH, —OR⁶, —N(R⁶)₂, or optionally        substituted alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl,        carbocyclylalkyl, heterocyclylalkyl, aralkyl, or        heteroarylalkyl, in which    -   each R⁶ is independently hydrogen, alkyl, carbocyclyl,        heterocyclyl, aryl, heteroaryl, carbocyclylalkyl,        heterocyclylalkyl, aralkyl, or heteroarylalkyl;    -   R⁴ is hydrogen, halogen, —OH, —OR⁶, —N(R⁶)₂, alkyl, carbocyclyl,        heterocyclyl, aryl, heteroaryl, carbocyclylalkyl,        heterocyclylalkyl, aralkyl, or heteroarylalkyl; and    -   R⁵ is hydrogen, methyl, ethyl, isoproplyl, t-butyl, CHF₂, CH₂F,        CF₃, CH₂OH, CHCH₃OH, or C(CH₃)₂OH.

In some embodiments, R² is CF₃.

At least one embodiment provides a pharmaceutical composition comprisinga compound of Formula I (wherein reference to Formula I includes FormulaIa and Ib and pharmaceutical salts thereof) or Formula II as describedherein. A pharmaceutical composition comprising a compound of Formula Ior Formula II may be used to treat cancer or other disease associatedwith abnormal histone demethylase activity. A pharmaceutical compositioncomprising a compound of Formula I or Formula II may be used in thepreparation of a medicament useful in treating or other diseaseassociated with abnormal histone demethylase activity.

One embodiment provides a method for inhibiting a histone demethylaseenzyme comprising contacting the histone demethylase enzyme with acompound of Formula I or Formula II. The contacting may be in vitro.

One embodiment provides a method of treating cancer or other diseaseassociated with abnormal histone demethylase activity in a subject inneed of such treatment, comprising administering to the subject apharmaceutical composition comprising a compound of Formula I or FormulaII as described herein, or a pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION

It should be understood that this invention is not limited to theparticular methodology, protocols, and reagents, etc., described hereinand as such may vary. The terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to limit thescope of the present invention, which is defined solely by the claims.

All patents and other publications identified are incorporated herein byreference for the purpose of describing and disclosing, for example, themethodologies described in such publications that might be used inconnection with the present invention, but are not to providedefinitions of terms inconsistent with those presented herein. Thesepublications are provided solely for their disclosure prior to thefiling date of the present application. Nothing in this regard should beconstrued as an admission that the inventors are not entitled toantedate such disclosure by virtue of prior invention or for any otherreason. All statements as to the date or representation as to thecontents of these documents is based on information available to theapplicants and do not constitute any admission as to the correctness ofthe dates or contents of these documents.

As used herein and in the claims, the singular forms “a,” “an,” and“the” include the plural reference unless the context clearly indicatesotherwise. Throughout this specification, unless otherwise indicated,“comprise,” “comprises” and “comprising” are used inclusively ratherthan exclusively, so that a stated integer or group of integers mayinclude one or more other non-stated integers or groups of integers. Theterm “or” is inclusive unless modified, for example, by “either.” Thus,unless context indicates otherwise, the word “or” means any one memberof a particular list and also includes any combination of members ofthat list. Other than in the operating examples, or where otherwiseindicated, all numbers expressing quantities of ingredients or reactionconditions used herein should be understood as modified in all instancesby the term “about.”

Headings are provided for convenience only and are not to be construedto limit the invention in any way. Unless defined otherwise, alltechnical and scientific terms used herein have the same meaning asthose commonly understood to one of ordinary skill in the art. Theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to limit the scope of the presentinvention, which is defined solely by the claims. Unless otherwisespecified, reference to a compound includes a pharmaceuticallyacceptable salt thereof. In other words, the term “compound” encompassespharmaceutically acceptable salts of that compound unless otherwisespecified by context. In order that the present disclosure can be morereadily understood, certain terms are defined. Additional definitionsare set forth throughout the detailed description.

Definitions

“Optional” or “optionally” means that a subsequently described event orcircumstance may or may not occur and that the description includesinstances when the event or circumstance occurs and instances in whichit does not. For example, “optionally substituted aryl” means that thearyl group may or may not be substituted and that the descriptionincludes both substituted aryl radicals and aryl radicals having nosubstitution. In a list of moieties, radical, or substitutents, the useof “optionally substituted” at the beginning of the list indicates andall member of the list are optionally substituted. In general, unlesscontext or language indicates otherwise, chemical groups or radicalsdescribed herein are optionally substituted.

“Alkyl” generally refers to a straight or branched hydrocarbon chainconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having generally from one to fifteen carbon atoms (e.g.,C₁-C₁₅ alkyl). In certain embodiments, an alkyl comprises one to eightcarbons (e.g., C₁-C₈ alkyl). In other embodiments, an alkyl comprisesone to five carbons (e.g., C₁-C₅ alkyl). In other embodiments, an alkylcomprises one carbon atom (e.g., C₁ alkyl) (e.g., methyl) or two carbons(e.g., C₂ alkyl) (e.g., ethyl). In other embodiments, an alkyl comprisesfive to fifteen carbon atoms (e.g., C₅-C₁₅ alkyl). In other embodiments,an alkyl comprises five to eight carbon atoms (e.g., C₅-C₈ alkyl). Inother embodiments, the alkyl group is selected from methyl, ethyl,1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl),1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl),1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl istypically attached to the rest of the molecule by a single bond. Unlessstated otherwise, an alkyl group is optionally substituted by at leastone substituent, such as halo, cyano, nitro, oxo, thioxo, imino, oximo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—OC(O)—N(R^(a))₂, —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a),—S(O)_(t)OR^(a), —S(O)_(t)R^(a or —S(O)) _(t)N(R^(a))₂, where t is 1 or2, where each R^(a) is independently hydrogen or optionally substitutedalkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocyclyl, heterocyclylalkyl, heteroaryl, or heteroarylalkyl, andwherein R^(a) is itself optionally substituted as described above. Forexample, in some embodiments, R^(a) is substituted with halogen,hydroxy, methoxy, or trifluoromethyl. These and other substituents areknown in the art. See, e.g., WO 2014089364, WO 2014100463, WO2014100818, WO 2014164708, WO 2014151945, WO 2014151106, WO 2015058160,WO 2015089192, WO 2015168466, WO 2015200709, WO 2015200843, WO2016004105, WO 2016003917, WO 2016037005, WO 2016044342, WO 2016044138,WO 2016044429, WO 2016168682, WO 2016172618.

“Alkoxy” refers to a radical bonded through an oxygen atom of theformula —O-alkyl, where alkyl is an alkyl chain as defined above; andunless stated otherwise, a moiety comprising an alkoxy group isoptionally substituted as described for alkyl.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon double bond, and having from two to twelvecarbon atoms. In certain embodiments, an alkenyl comprises two to eightcarbon atoms. In other embodiments, an alkenyl comprises two to fourcarbon atoms. The alkenyl is attached to the rest of the molecule by asingle bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e.,allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. Unlessstated otherwise specifically in the specification, an alkenyl group isoptionally substituted as described for alkyl.

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon triple bond, having from two to twelve carbonatoms. In certain embodiments, an alkynyl comprises two to eight carbonatoms. In other embodiments, an alkynyl has two to four carbon atoms.The alkynyl is attached to the rest of the molecule by a single bond,for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and thelike. Unless stated otherwise, an alkynyl group is optionallysubstituted as described for alkyl.

“Alkylene” “alkylene chain” “alkyl chain” or “alkyl linker” refers to astraight or branched divalent hydrocarbon chain linking the rest of themolecule to a radical group, and consisting solely of carbon andhydrogen, containing no unsaturation and having from one to twelvecarbon atoms, for example, methylene, ethylene, propylene, n-butylene,and the like. Reference to alkyl may refer to such chains or linkers, asindicated by context. Similarly, “alkynylene chain” refers to a straightor branched divalent hydrocarbon chain linking the rest of the moleculeto a radical group, consisting solely of carbon and hydrogen, containingat least one carbon-carbon triple bond and having from two to twelvecarbon atoms. These hydrocarbon chains are optionally substituted asdescribed for alkyl.

“Aryl” refers to an aromatic monocyclic or multicyclic hydrocarbon ringsystem containing a delocalized (4n+2) π-electron system in accordancewith the Hückel theory. The aryl contains only hydrogen and carbon,generally from five to eighteen carbon atoms, where at least one of therings in the ring system is fully unsaturated. Aryls include benzene,fluorene, indane, indene, tetralin, and naphthalene. Unless statedotherwise, the term “aryl” or the prefix “ar-” (such as in “aralkyl”)includes aryls optionally substituted by one or more substituentsindependently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)C(O)N(R^(a))₂,—R^(b)—O—R^(c)—C(O)N(R^(a))₂, —R^(b)—N(R^(a))C(O)OR^(a),—R^(b)—N(R^(a))C(O)R^(a), —R^(b)—N(R^(a))S(O)_(t)R^(a),—R^(b)—S(O)_(t)OR^(a), —R^(b)—S(O)_(t)R^(a), or—R^(b)—S(O)_(t)N(R^(a))₂, where t is 1 or 2, where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl (optionally substituted with one or more halo groups), aralkyl,heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, eachR^(b) is independently a direct bond or a straight or branched alkyleneor alkenylene chain, and R^(c) is a straight or branched alkyl oralkenylene chain, and where each of the above substituents is optionallysubstituted unless otherwise indicated. Additional substituents are asdescribed for alkly.

“Aralkyl” refers to the formula —R^(c)-aryl where R^(c) is an alkyl oralkyl chain (e.g., alkyl linker) as defined above, for example,methylene, ethylene, and the like. The alkyl chain part of the aralkylis optionally substituted as described above for alkyl. The aryl part ofthe aralkyl radical is optionally substituted as described for an arylgroup.

“Aralkenyl” refers to a group with the formula —R^(d)-aryl where R^(d)is an alkenylene chain as defined above. The aryl part of the aralkenylradical is optionally substituted as described above for an aryl group.The alkenylene chain part of the aralkenyl radical is optionallysubstituted as defined above for an alkenylene group.

“Aralkynyl” refers to a radical of the formula Rearyl, where Re is analkynylene chain as defined above. The aryl part of the aralkynylradical is optionally substituted as described above for an aryl group.The alkynylene chain part of the aralkynyl radical is optionallysubstituted as defined above for an alkynylene chain.

“Aralkoxy” refers to a group bonded through an oxygen atom of theformula —O—R^(c)-aryl where R^(c) is an alkyl chain as defined above,for example, methylene, ethylene, and the like. The alkyl chain part ofthe aralkyl is optionally substituted as described above for an alkylchain. The aryl part of the aralkyloxy is optionally substituted asdescribed above for an aryl group.

“Carbocyclyl” refers to a stable non-aromatic (saturated) monocyclic,bicyclic, or polycyclic hydrocarbon group consisting solely of carbonand hydrogen atoms, which generally includes fused or bridged ringsystems, having from three to fifteen carbon atoms. In certainembodiments, a carbocyclyl comprises three to ten carbon atoms. In otherembodiments, a carbocyclyl comprises three to seven carbon atoms. Thecarbocyclyl is attached to the rest of the molecule by single bond(s). Acarbocyclyl group may be fully saturated or partially saturated. A fullysaturated carbocyclyl group may also refer to a “cycloalkyl.” Examplemonocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl mayalso refer to a “cycloalkenyl.” Example monocyclic cycloalkenyls includecyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycycliccarbocyclyls include, for example, adamantyl, norbornyl (i.e.,bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl,7,7-dimethyl-bicyclo[2.2.1]-heptanyl, and the like. Unless otherwisestated, the term “carbocyclyl” includes carbocyclyls that are optionallysubstituted by one or more substituents independently selected and asdescribed above.

“Carbocyclylalkyl” refers to a group of the formula —R^(c)-carbocyclyl,wherein R^(c) is an alkyl chain, optionally substituted, as describedabove. Similarly, Carbocyclylalkynyl” refers to a group of the formula—R^(c)-carbocyclyl, where R^(c) is an alkynylene chain, optionallysubstituted, as defined above. In some embodiments the carbocyclyl groupis a cycloalkyl group, in which the alkynylene chain part of thecarbocyclylalkynyl is optionally substituted as defined above for analkyl chain.

“Carbocyclylalkoxy” refers to a radical bonded through an oxygen atom ofthe formula —O—R^(c)-carbocyclyl where R^(c) is an alkyl chain,optionally substituted, as defined above for alkyl.

As used herein, “carboxylic acid bioisostere” refers to a functionalgroup or moiety that exhibits similar physical, biological and/orchemical properties as a carboxylic acid moiety. Examples of carboxylicacid bioisosteres include, but are not limited to,

and the like.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodosubstituents.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, fluoromethyl,2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. Thealkyl part of the fluoroalkyl radical may be optionally substituted asdefined above for alkyl.

“Heterocyclyl” refers to a stable 3- to 18-membered non-aromatic ringradical that comprises two to twelve carbon atoms and from one to sixheteroatoms selected from nitrogen, oxygen and sulfur. Unless statedotherwise specifically in the specification, the heterocyclyl radical isa monocyclic, bicyclic, tricyclic or tetracyclic ring system, which mayinclude fused or bridged ring systems. The heteroatoms in theheterocyclyl radical may be optionally oxidized. One or more nitrogenatoms, if present, are optionally quaternized. The heterocyclyl radicalis partially or fully saturated. The heterocyclyl may be attached to therest of the molecule through any atom of the ring(s). Examples of suchheterocyclyl radicals include, but are not limited to, azocanyl,chromenyl, cinnolinyl, dioxolanyl, thienyl[1,3]dithianyl,decahydroquinolyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl,isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl,octahydroisoindolyl, 2-oxopiperazinyl, 2-oxo-piperidinyl,2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl,4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl,thiocanyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thio-morpholinyl. Unless stated otherwise, the term“heterocyclyl” includes heterocyclyl group as defined above that areoptionally substituted by one or more substituents as described herein.

“N-heterocyclyl” or “N-attached heterocyclyl” refers to a heterocyclylradical as defined above containing at least one nitrogen and where thepoint of attachment of the heterocyclyl radical to the rest of themolecule is through a nitrogen atom in the heterocyclyl radical. AnN-heterocyclyl radical is optionally substituted as described above forheterocyclyl radicals. Examples of such N-heterocyclyl radicals include,but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl,1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

“C-heterocyclyl” or “C-attached heterocyclyl” refers to a heterocyclylradical as defined above containing at least one heteroatom and wherethe point of attachment of the heterocyclyl radical to the rest of themolecule is through a carbon atom in the heterocyclyl radical. AC-heterocyclyl radical is optionally substituted as described above forheterocyclyl radicals. Examples of such C-heterocyclyl radicals include,but are not limited to, 2-morpholinyl, 2- or 3- or 4-piperidinyl,2-piperazinyl, 2- or 3-pyrrolidinyl, and the like.

“Heterocyclylalkyl” refers to a radical of the formula—R^(c)-heterocyclyl where R^(c) is an alkylene chain as defined above.If the heterocyclyl is a nitrogen-containing heterocyclyl, theheterocyclyl is optionally attached to the alkyl radical at the nitrogenatom. The alkyl chain of the heterocyclylalkyl and the heterocyclyl partof the heterocyclylalkyl group may each be optionally substituted asdefined above.

“Heterocyclylalkoxy” refers to a radical bonded through an oxygen atomof the formula —O—R^(c)-heterocyclyl where R^(c) is an alkylene chain asdefined above. If the heterocyclyl is a nitrogen-containingheterocyclyl, the heterocyclyl is optionally attached to the alkylradical at the nitrogen atom. The alkyl chain of the heterocyclylalkoxyis optionally substituted as defined above for alkyl. The heterocyclylpart of the heterocyclylalkoxy is optionally substituted as definedabove for a heterocyclyl group.

Heteroaryl” refers to a moiety derived from a three- toeighteen-membered aromatic ring that generally comprises two toseventeen carbon atoms and from one to six heteroatoms selected fromnitrogen, oxygen and sulfur, wherein at least one of the rings in thering system is fully unsaturated, i.e., it contains a cyclic,delocalized (4n+2) π-electron system in accordance with the Hückeltheory. The heteroaryl may be a monocyclic, bicyclic, tricyclic ortetracyclic ring system, and includes fused or bridged ring systems. Theheteroatom(s) in the heteroaryl group is optionally oxidized. One ormore nitrogen atoms, if present, are optionally quaternized. Theheteroaryl can be attached to the rest of the molecule through any atomof the ring(s). Unless stated otherwise, heteroaryls are optionallysubstituted with one or more substituents as described herein.

Examples of heteroaryls include, but are not limited to, azepinyl,acridinyl, benzimidazolyl, benzin-dolyl, 1,3-benzodioxolyl,benzofuranyl, benzooxazolyl, benzo[d]-thiazolyl, benzothiadiazolyl,benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzo-dioxanyl,benzonaphthofuranyl, benzo-xazolyl, benzodioxolyl, benzodioxinyl,benzopyranyl, benzopyranonyl, benzofuranyl, benzo-furanonyl,benzothienyl (benzothiophenyl), benzo-thieno[3,2-d]pyrimidinyl,benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl,cinnolinyl, cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo [h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]-cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzo-thiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d] pyrimidinyl,5,6,7,8,9,10-hexahydro-cycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d] pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquina-zolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7, 8,9,10,10a-octa-hydrobenzo[h]quin-azolinyl,1-phenyl-1H-pyrrolyl, phenazinyl, pheno-thiazinyl, phenoxazinyl,phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]-pyrimidinyl,pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl,quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl,tetrahydroquinolinyl, 5,6,7,8-tetrahydro-quinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclo-hepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c] pyridazinyl, thiazolyl, thiadiazolyl,triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimi-dinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl).

“N-heteroaryl” refers to a heteroaryl containing at least one nitrogenand where the point of attachment of the heteroaryl to the rest of themolecule is through a nitrogen atom in the heteroaryl radical. AN-heteroaryl is optionally substituted as described above.

“C-heteroaryl” refers to a heteroaryl where the point of attachment ofthe heteroaryl to the rest of the molecule is through a carbon atom inthe heteroaryl. A C-heteroaryl is optionally substituted as describedabove for heteroaryl.

“Heteroarylalkyl” refers to a group of the formula —R^(c)-heteroaryl,where R^(c) is an alkylene chain as defined above. If the heteroaryl isa nitrogen-containing heteroaryl, the heteroaryl is optionally attachedto the alkyl radical at the nitrogen atom. The alkylene chain of theheteroarylalkyl radical is optionally substituted as defined above foran alkylene chain. The heteroaryl part of the heteroarylalkyl radical isoptionally substituted as defined above for a heteroaryl group.

“Heteroarylalkoxy” refers to a radical bonded through an oxygen atom ofthe formula —O—R^(c)-heteroaryl, where R^(c) is an alkyl chain. If theheteroaryl is a nitrogen-containing heteroaryl, the heteroaryl isoptionally attached to the alkyl radical at the nitrogen atom. The alkylchain of the heteroarylalkoxy is optionally substituted as defined abovefor alkyl. The heteroaryl part of the heteroarylalkoxy is optionallysubstituted as defined above for heteroaryl.

The compounds disclosed herein may contain one or more asymmetriccenters and may thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)— or (S)—. Unless stated otherwise, it isintended that all stereoisomeric forms of the compounds disclosed hereinare contemplated by this disclosure. When the compounds described hereincontain alkene double bonds, and unless specified otherwise, it isintended that this disclosure includes both E and Z geometric isomers(e.g., cis or trans). Likewise, all possible isomers, as well as theirracemic and optically pure forms, and all tautomeric forms are alsointended to be included. The term “geometric isomer” refers to E or Zgeometric isomers (e.g., cis or trans) of an alkene double bond. Theterm “positional isomer” refers to structural isomers around a centralring, such as ortho-, meta-, and para-isomers around a benzene ring.

A “tautomer” refers to a molecule wherein a proton shift from one atomof a molecule to another atom of the same molecule is possible. Thecompounds presented herein may, in certain embodiments, exist astautomers. In circumstances where tautomerization is possible, achemical equilibrium of the tautomers will exist. The exact ratio of thetautomers depends on several factors, including physical state,temperature, solvent, and pH. Some examples of tautomeric equilibriuminclude:

“Pharmaceutically acceptable salt” includes both acid and base additionsalts. A pharmaceutically acceptable salt of any one of the compoundsdescribed herein is intended to encompass any and all pharmaceuticallysuitable salt forms. A pharmaceutically acceptable salt of any one ofthese compounds is intended to encompass any and all pharmaceuticallysuitable salt forms, including pharmaceutically acceptable salts such asacid and base addition salts, as are well-known in the art. See, e.g.,WO 2014089364, WO 2014100463, WO 2014100818, WO 2014164708, WO2014151945, WO 2014151106, WO 2015058160, WO 2015089192, WO 2015168466,WO 2015200709, WO 2015200843, WO 2016004105, WO 2016003917, WO2016037005, WO 2016044342, WO 2016044138, WO 2016044429, WO 2016168682,WO 2016172618.

Further, a compound described herein may be produced or formulated as a“prodrug.” Prodrugs are compounds that may be inactive whenadministered, but are converted under physiological conditions or byhydrolysis (i.e., in vivo) to a biologically active compound; thusprodrugs are pharmaceutically acceptable precursors of a biologicallyactive compound. Prodrug compounds may offer advantages of solubility,tissue compatibility, or delayed release in a subject. Prodrugs alsorefer to use of covalently bonded carriers that release the activecompound in vivo when such prodrug is administered to the subject.Prodrugs of an active compound may be prepared by modifying functionalgroups present in the active compound in such a way that themodifications are cleaved, either in routine manipulation or in vivo, tothe parent active compound. For example, prodrugs include compounds inwhich a hydroxy, amino, or mercapto group is bonded to any group that,when the prodrug of the active compound is administered to a mammaliansubject, cleaves to form a free hydroxy, free amino, or free mercaptogroup, respectively. Examples of prodrugs include acetate, carboxylate,formate, and benzoate derivatives of alcohol or amine functional groupsin the active compounds. See, e.g., Bundgard, DESIGN OF PRODRUGS, at7-9, 21-24 (Elsevier, Amsterdam, 1985); Higuchi et al., Pro drugs asNovel Delivery Systems, 14 A.C.S. Symposium Series; BIOREVERSIBLECARRIERS IN DRUG DESIGN (Edward B. Roche (Ed.), Am. Pharm. Assoc. andPergamon Press, 1987).

Accordingly, and as used herein, reference to “compound” or “Formula” or“compound of Formula I,” and the like, includes within that reference apharmaceutically acceptable salt, hydrate, solvate, N-oxide,stereoisomer, tautomer, radioisotopically enriched or deuteratedversion, or prodrug thereof.

As used herein, “treatment” or “treating,” or “palliating” or“ameliorating” are used interchangeably herein. These terms refer to anapproach for obtaining beneficial or desired results including but notlimited to therapeutic benefit or a prophylactic benefit. By“therapeutic benefit” is meant eradication or amelioration of theunderlying disorder being treated. Also, a therapeutic benefit isachieved with the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient, notwithstanding that thepatient may still be afflicted with the underlying disorder. Forprophylactic benefit, the compositions may be administered to a patientat risk of developing a particular disease, or to a patient reportingone or more of the physiological symptoms of a disease, even though adiagnosis of this disease may not have been made.

The term “modulating” means, in relation to histone demethylation, aprocess in which the enzymatic activity of the histone demethylase ischanged by contact with a substituted triazolylpyridine derivativecompound described herein in comparison with a control; the activitylevel may be increased or decreased (i.e., inhibited) depending on theparticular cellular or molecular context of the histone demethylase. Theoriginal, unmodulated activity may be of any kind of activity, includingabsence of any activity. The term “modulating the activity” includes,for example, any change of histone demethylase activity that leads toincreased function of a cellular pathway (e.g., a signaling pathway),including pathways that conclude with apoptosis. The enzymatic activitycan increase from zero (absent or immeasurable activity) to a certainamount; or, more typically, can decrease from a certain amount to andimmeasurable small amount or zero. Modulating activity can be expressed,for example, as IC₅₀ when a substituted triazolylpyridine derivativecompound inhibits enzymatic activity of a histone demethylase (seeExamples).

Substituted Triazolylpyridine Derivative Compounds

Substituted compounds are described herein that inhibit a histonedemethylase enzyme. These compounds, and compositions comprising thesecompounds, are useful for the treatment of cancer and neoplasticdiseases. The compounds described herein may, therefore, be useful fortreating pancreatic cancer, prostate cancer, breast cancer, bladdercancer, lung cancer, gastric cancer, leukemia or melanoma and the like.As noted above: unless stated otherwise all references to a compoundinclude the pharmaceutically acceptable salts thereof.

At least one embodiment provides a compound having the structure ofFormula I

wherein the compound of Formula I includes pharmaceutical salts thereof,and wherein

-   -   R¹ is halogen, CH₂G, NHG, or OG, wherein        -   G is hydrogen, alkyl, or —X—Y, in which            -   X is hydrogen or —C₁ alkylene, and            -   Y is optionally substituted aralkyl, aralkenyl,                aralkynyl, carbocyclyl, carbocyclylalkyl, heterocyclyl,                heterocyclylalkyl, heteroaryl, heteroarylalkyl,                adamantyl, benzofuranyl, 2,3-dihydrobenzofuranyl,                chromanyl, indanyl, indolyl, naphthyl, optionally                substituted 1,2-dihydronaphthyl, phenyl, pyridyl,                tetrahydroquinolinyl, tetralinyl,                2,3-dihydrobenzo[b][1,4]dioxinyl or thiochromanyl, or                —R^(b)—OR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a),                —R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))₂,                —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),                —R^(b)-OvR^(c)—C(O)N(R^(a))₂, —R^(b)—N(R^(a))C(O)OR^(a),                —R^(b)—N(R^(a))C(O)R^(a), —R^(b)—N(R^(a))S(O)_(t)R^(a),                —R^(b)—S(O)_(t)OR^(a), —R^(b)—S(O)_(t)OR^(a), or                —R^(b)—S(O)_(t)N(R^(a))₂, wherein                -   each R^(a) is independently hydrogen, alkyl,                    fluoroalkyl, cycloalkyl, cycloalkylalkyl, aryl,                    aralkyl, heterocyclyl, heterocyclylalkyl,                    heteroaryl, or heteroarylalkyl,                -   each R^(b) is independently a direct bond or a                    straight or branched alkylene or alkenylene chain,                -   each R^(c) is a straight or branched alkylene or                    alkenylene chain; and                -   t is 1 or 2;    -   R² is halogen or CF₃; and    -   R⁵ hydrogen, methyl, ethyl, isoproplyl, t-butyl, CHF₂, CH₂F,        CF₃, CH₂OH, CHCH₃OH, or C(CH₃)₂OH.

In some embodiments of the compound of Formula I, R¹ is fluoro.

In some embodiments of the compound of Formula I, R² is CF₃.

In some embodiments of the compound of Formula I, R⁵ is hydrogen.

In some embodiments of the compound of Formula I, Y is phenyl optionallysubstituted with alkl, alkynyl, chloro, fluoro, fluoroalkyl, nitro oroptionally substituted aralkyl, aralkenyl, aralkynyl, carbocyclyl,carbocyclylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl,heteroarylalkyl, or —R^(b)—OR^(a), —R^(b)—OC(O)—R^(a),—R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))₂,—R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a), —R^(b)-OvR^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a), —R^(b)—S(O)_(t)OR^(a),—R^(b)—S(O)_(t)OR^(a), or —R^(b)—S(O)_(t)N(R^(a))₂, in which each R^(a)is independently hydrogen, alkyl, fluoroalkyl, cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl, or heteroarylalkyl; each R^(b) is independently a directbond or a straight or branched alkyl or alkenyl chain; each R^(c) is astraight or branched alkyl or alkenyl chain; and t is 1 or 2.

Another embodiment provides a compound of Formula I wherein R¹ is alkyl.Another embodiment provides a compound of Formula I, wherein R¹ ismethyl.

Another embodiment provides a compound of Formula I, wherein R¹ iscarbocyclyl or carbocyclylalkyl. Another embodiment provides a compoundof Formula I, wherein R¹ is carbocyclyl(C₁-C₆ alkyl). Another embodimentprovides a compound of Formula I, wherein R¹ is carbocyclylethyl orcarbocyclylmethyl.

Another embodiment provides a compound of Formula I, wherein R¹ iscarbocyclyl(C₁-C₆ alkyl) and the carbocyclyl is1,2,3,4-tetrahydronaphthyl optionally substituted with one or moregroups selected from halogen, hydroxy, —CN, alkyl, alkoxy, alkylamino,aryl, carbocyclyl, heterocyclyl, carbocyclylalkyl, or heterocyclylalkyl.

Another embodiment provides a compound of Formula I wherein R¹ isoptionally substituted heterocyclyl or heterocyclylalkyl.

Another embodiment provides a compound of Formula I wherein R¹ isoptionally substituted heteroaryl or heteroarylalkyl.

Another embodiment provides a compound of Formula I wherein R¹ isoptionally substituted aryl or aralkyl.

Another embodiment provides a compound of Formula I wherein R¹ isoptionally substituted aralkyl, and the aralkyl is aryl(C₁-C₆ alkyl).Another embodiment provides a compound of Formula I wherein R¹ isoptionally substituted aralkyl, the aralkyl is arylethyl or arylmethyl.

Another embodiment provides a compound of Formula I, wherein Y isoptionally substituted aryl or aralkyl, and the aryl is phenyloptionally substituted with at least one halogen, hydroxy, —CN, alkyl,alkoxy, alkylamino, aryl, carbocyclyl, heterocyclyl, carbocyclylalkyl,or heterocyclylalkyl.

Another embodiment provides a compound of Formula I, wherein Y isoptionally substituted aryl or aralkyl, and the aryl is naphthyloptionally substituted with at least one halogen, hydroxy, —CN, alkyl,alkoxy, alkylamino, aryl, carbocyclyl, heterocyclyl, carbocyclylalkyl,or heterocyclylalkyl.

Another embodiment provides a compound of Formula I wherein Y ishydrogen, halogen, —CN, or an alkyl optionally substituted with at leastone fluoro. Another embodiment provides a compound of Formula I whereinX is hydrogen. Another embodiment provides a compound of Formula Iwherein R¹ is fluoro. Another embodiment provides a compound of FormulaI wherein R¹ is chloro. Another embodiment provides a compound ofFormula I wherein R¹ is iodo. Another embodiment provides a compound ofFormula I wherein Y is —CN. Another embodiment provides a compound ofFormula I wherein R² is —CF₃.

Another embodiment provides a compound of Formula I wherein R¹ ishydrogen or aryl. Another embodiment provides a compound of Formula Iwherein R¹ is hydrogen. Another embodiment provides a compound ofFormula I wherein R¹ is aryl. Another embodiment provides a compound ofFormula I wherein R¹ is aryl and the aryl is phenyl optionallysubstituted with at least one halogen, hydroxy, —CN, alkyl, alkoxy,—O-(cycloalkylalkyl), alkylamino, aryl, carbocyclyl, heterocyclyl,carbocyclylalkyl, or heterocyclylalkyl. Another embodiment provides acompound of Formula I wherein R¹ is aryl, and the aryl is a phenyl issubstituted with one or more groups selected from halogen, alkoxy, or—O-(cycloalkylalkyl). Another embodiment provides a compound of FormulaI wherein R⁵ is hydrogen.

In at least one embodiment, the compound of Formula I has the structure:

wherein G, R², and R⁵ are as described above.

In at least one embodiment, the compound of Formula I has the structure:

-   -   wherein    -   R² and R⁵ are as described above; and    -   Z is independently at least one hydrogen, halogen, —OH, —CN, or        optionally substituted C₁-C₆ alkyl, C₁-C₆ alkoxy, C₃-C₇        carbocyclyl, C₃-C₇ carbocyclyloxy, C₄-C₁₂ carbocyclylalkyl,        C₄-C₁₂ carbocyclylalkoxy, C₁-C₆ alkynyl, C₁-C₆ alkenyl, C₆-C₁₀        aryl, C₆-C₁₀ aryloxy, C₆-C₁₀ aryl-S—, C₇-C₁₄ aralkoxy,        heteroaryl, or heteroaryloxy.

In some embodiments of a compound of Formula Ib, Z is at least onehalogen. In some embodiments of the compound, Z is at least one fluoro.In some embodiments, R² is CF₃. In some embodiments, R⁵ is hydrogen.

In another embodiment of a compound of Formula Ib, Z is independently atleast one hydroxy, halogen, cyano, NH₂, NHR^(d), N(R^(d))₂, NHC(O)R^(d),NHC(O)OR^(d), NHC(O)NHR^(d), NHC(O)N(R^(d))₂, NHS(O)₂R^(d),NR^(d)C(O)R^(d), NR^(d)C(O)OR^(d), NR^(d)C(O)NHR^(d),NR^(d)C(O)N(R^(d))₂, NR^(d)S(O)₂R^(d), or optionally substituted alkyl,alkenyl, alkynyl, alkoxy, aryl, aryloxy, aralkyl, carbocyclyl,heterocyclyl, heteroaryl, carbocyclylalkyl, heterocyclylalkyl, orheteroarylalkyl; in which each R^(d) is independently alkyl, aryl,aralkyl, carbocyclyl, heterocyclyl, heteroaryl, carbocyclylalkyl,heterocyclylalkyl or heteroarylalkyl.

In some embodiments of a compound of Formula Ib, R² is CF₃, one of Z isfluoro; one of Z is an optionally substituted straight, branched, orcyclic C₁-C₆ alkyl, and R⁵ is hydrogen. In some embodiments of thecompound, R² is CF₃, one of Z is fluoro and one of Z is an optionallysubstituted straight, branched, or cyclic C₁-C₆ alkoxy, and R⁵ ishydrogen. In some embodiments of the compound, R² is CF₃, one of Z isfluoro, one of Z is trifluoromethyloxy, and R⁵ is hydrogen. In someembodiments of the compound of Formula I, R² is CF₃, one of Z is fluoroand one of Z is phenylmethoxy, and R⁵ is hydrogen. In some embodimentsof the compound, R² is CF₃, one of Z is fluoro, one of Z is NHR^(d) orN(R^(d))₂, and R⁵ is hydrogen. In some embodiments of the compound, R²is CF₃, R¹ is NHG, wherein G is —X—Y and X is —C₁ alkylene, and R⁵ ishydrogen.

At least one embodiment provides a compound having the structure ofFormula II:

-   -   wherein a compound of Formula II includes pharmaceutically        acceptable salts thereof, and wherein        -   R² is halogen or CF₃;        -   R³ and R⁴ are each independently hydrogen, halogen, —OH,            —OR⁶, —N(R⁶)₂, alkyl, carbocyclyl, heterocyclyl, aryl,            heteroaryl, carbocyclylalkyl, heterocyclylalkyl, aralkyl, or            heteroarylalkyl, in which        -   each R⁶ is independently hydrogen, alkyl, carbocyclyl,            heterocyclyl, aryl, heteroaryl, carbocyclylalkyl,            heterocyclylalkyl, aralkyl, or heteroarylalkyl; and        -   R⁵ is hydrogen, methyl, ethyl, isoproplyl, t-butyl, —CHF₂,            —CH₂F, —CF₃, —CH₂OH, —CHCH₃OH, or —C(CH₃)₂OH.

Some embodiments provide a compound of Formula II in which R² is CF₃.Some embodiments provide a compound of Formula II wherein R⁵ ishydrogen.

Other embodiments provide a compound of Formula II wherein R⁴ isoptionally substituted C₁-C₄ alkyl. Another embodiment provides acompound of Formula II wherein R⁴ is C₁-C₄ alkyl, and the alkyl issubstituted with at least one fluoro. Another embodiment provides acompound of Formula II wherein R⁴ is CH₂F, CHF₂, or CF₃. Anotherembodiment provides a compound of Formula II wherein R⁴ is CF₃. Anotherembodiment provides a compound of Formula II wherein R⁵ is H.

Another embodiment provides the compound of Formula II wherein R⁴ isaryl. Another embodiment provides the compound of Formula II, wherein R⁴is heteroaryl. Another embodiment provides the compound of Formula IIwherein R⁴ is aryl, and each aryl is independently optionallysubstituted with halogen, alkoxy, carbocyclyloxy, heterocyclyloxy,aryloxy, heteroaryloxy, carbocyclylalkoxy, heterocyclylalkoxy,aralkyloxy, or heteroarylalkoxy. Another embodiment provides thecompound of Formula II wherein R⁴ is aryl, and the aryl is optionallysubstituted with halogen, alkoxy, carbocyclyloxy heterocyclyloxy,aryloxy, heteroaryloxy, carbocyclylalkoxy, heterocyclylalkoxy, aralkoxy,or heteroarylalkoxy.

Another embodiment provides a compound of Formula II wherein R³ ishydrogen. Another embodiment provides a compound of Formula II whereinR³ is alkyl. Another embodiment provides a compound of Formula IIwherein R² is methyl.

Another embodiment provides a compound of Formula II, wherein R³ isoptionally substituted carbocyclyl or carbocyclylalkyl. Anotherembodiment provides a compound of Formula II, or a pharmaceuticallyacceptable salt thereof, wherein R³ is carbocyclyl(C₁-C₆ alkyl). Anotherembodiment provides a compound of Formula II wherein R³iscarbocyclyl(C₁-C₆ alkyl), in which the C₁-C₆ alkyl is ethyl or methyl.

Another embodiment provides a compound of Formula II, wherein R³ isoptionally substituted carbocyclyl(C₁-C₆ alkyl), and the carbocyclyl is1,2,3,4-tetrahydronaphthyl optionally substituted with at least onehalogen, hydroxy, —CN, alkyl, alkoxy, cycloalkylalkoxy, alkylamino,aryl, carbocyclyl, heterocyclyl, carbocyclylalkyl, or heterocyclylalkyl.

Another embodiment provides a compound of Formula II, or apharmaceutically acceptable salt thereof, wherein R³ is optionallysubstituted heterocyclyl or heterocyclylalkyl. Another embodimentprovides a compound of Formula II, or a pharmaceutically acceptable saltthereof, wherein R³ is optionally substituted heterocyclyl(C₁-C₆ alkyl).

Another embodiment provides a compound of Formula II wherein R³ isoptionally substituted heteroaryl or heteroarylalkyl. Another embodimentprovides a compound of Formula II wherein R³ is optionally substitutedheterocyclylalkyl, and R⁴ and R⁵ are hydrogen. Another embodimentprovides a compound of Formula II wherein R³ is optionally substitutedcyclylalkyl, and R⁴ and R⁵ are hydrogen. Another embodiment provides acompound of Formula II wherein R³ is phenyloxymethyl and R⁴ and R⁵ arehydrogen.

Another embodiment provides a compound of Formula II, wherein R³ isoptionally substituted heteroaryl or heteroarylalkyl, and the heteroarylis pyridine or pyrimidine optionally substituted with at least onehalogen, hydroxy, —CN, alkyl, alkoxy, cycloalkylalkoxy, alkamino, aryl,carbocyclyl, heterocyclyl, carbocyclylalkyl, or heterocyclylalkyl.

Another embodiment provides a compound of Formula II, wherein R³ isoptionally substituted heteroaryl, and the heteroaryl is a chromanyloptionally substituted with at least one halogen, hydroxy, —CN, alkyl,alkoxy, cycloalkylalkoxy, alkylamino, aryl, carbocyclyl, heterocyclyl,carbocyclylalkyl, or heterocyclylalkyl.

Another embodiment provides a compound of Formula II wherein R³ isoptionally substituted heteroarylalkyl, and the heteroarylalkylcomprises a chromanyl optionally substituted with at least one halogen,hydroxy, —CN, alkyl, alkoxy, cycloalkylalkoxy, alkylamino, aryl,carbocyclyl, heterocyclyl, carbocyclylalkyl, or heterocyclylalkyl.Another embodiment provides a compound of Formula II wherein R³ is ethylor methyl. Another embodiment provides a compound of Formula II whereinR³ is aryl or aralkyl. Another embodiment provides the compound ofFormula II wherein R⁵ is hydrogen.

Another embodiment provides a compound of Formula II wherein R³ isoptionally substituted aryl or aralkyl, and the aryl is a phenyloptionally substituted with at least one halogen, hydroxy, —CN, alkyl,alkoxy, cycloalkylalkoxy, alkamino, aryl, carbocyclyl, heterocyclyl,carbocyclylalkyl, or heterocyclylalkyl. Another embodiment provides acompound of Formula II wherein R² is aryl or aralkyl, and the aryl isphenyl optionally substituted with at least one halogen, alkoxy, oralkyl.

Another embodiment provides a compound of Formula II wherein R³ isoptionally substituted aralkyl, and the aralkyl is aryl(C₁-C₆ alkyl).Another embodiment provides a compound of Formula II wherein R³ isaralkyl, the aralkyl is aryl(C₁-C₆ alkyl), and the aryl is phenyloptionally substituted with one or more of halogen, hydroxy, —CN, alkyl,alkoxy, cycloalkylalkoxy, alkylamino, aryl, carbocyclyl, heterocyclyl,carbocyclylalkyl, or heterocyclylalkyl. Another embodiment provides acompound of Formula II wherein R³ is aralkyl, the aralkyl is aryl(C₁-C₆alkyl), and the (C₁-C₆ alkyl) is ethyl or methyl.

Another embodiment provides a compound of Formula II wherein R³ isaralkyl, and the aralkyl comprises a naphthyl optionally substitutedwith at least one halogen, hydroxy, —CN, alkyl, alkoxy,cycloalkylalkoxy, alkylamino, aryl, carbocyclyl, heterocyclyl,carbocyclylalkyl, or heterocyclylalkyl. Another embodiment provides acompound of Formula II wherein the aralkyl further comprises a C₁alkylene, or a C₂ alkylene.

Another embodiment provides a compound of Formula II, wherein R¹ or R⁵are hydrogen. Another embodiment provides a compound of Formula II,wherein both R¹ and R⁵ are hydrogen.

Another embodiment provides a compound of Formula II wherein R³ is aryl.Another embodiment provides a compound of Formula II wherein R³ ishydrogen or aryl.

Another embodiment provides a compound of Formula II, or apharmaceutically acceptable salt thereof, wherein R³ is aryl, and thearyl is a phenyl optionally substituted with one or more groups selectedfrom halogen, hydroxy, —CN, alkyl, alkoxy, cycloalkylalkoxy, alkylamino,aryl, carbocyclyl, heterocyclyl, carbocyclylalkyl, or heterocyclylalkyl.Another embodiment provides a compound of Formula II, or apharmaceutically acceptable salt thereof, wherein R¹ is aryl, and thearyl is a phenyl optionally substituted with one or more groups selectedfrom halogen, alkoxy, or cycloalkylalkoxy.

In some embodiments, a compound as disclosed herein has the structureprovided in Table 1:

TABLE 1 Chemical Synthesis Example Structure Name 13-fluoro-4-[5-(trifluoromethyl)-1H-1,2,3- triazol-4-yl]pyridine 2N-(5-chloro-2-fluorophenyl)-4-[5- (trifluoromethyl)-1H-1,2,3-triazol-4-yl]pyridin-3-amine 3 N-[(1,2,3,4-tetrahydronaphthalen-1-yl)methyl]-4-[5-(trifluoromethyl)-1H-1,2,3- triazol-4-yl]pyridin-3-amine4 3-[(4-chlorophenyl)methyl]-1-{4-[5-(trifluoromethyl)-1H-1,2,3-triazol-4- yl]pyridin-2-yl}-1H-pyrazol-5-ol

In some embodiments, the compound disclosed herein has the structureprovided in Table 2, where R is Cl, F, or CF₃.

TABLE 2

Preparation of the Substituted Triazolylpyridine Derivative Compounds

The compounds used in the reactions described herein are made accordingto organic synthesis techniques known to those skilled in this art,starting from commercially available chemicals and/or from compoundsdescribed in the chemical literature. “Commercially available chemicals”are obtained from standard commercial sources including Acros Organics(Pittsburgh, Pa., US), Aldrich Chemical (Milwaukee, Wis., US; includesSigma Chemical and Fluka), Apin Chemicals Ltd. (Milton Park, UK),Avocado Research (Lancashire, UK), BDH Inc. (Toronto, CA), Bionet(Cornwall, UK), Chemservice Inc. (West Chester, Pa., US), CrescentChemical Co. (Hauppauge, N.Y., US), Eastman Organic Chemicals, EastmanKodak Company (Rochester, N.Y., US), Fisher Scientific Co. (Pittsburgh,Pa., US), Fisons Chemicals (Leicestershire, UK), Frontier Scientific(Logan, Utah, US), ICN Biomedicals, Inc. (Costa Mesa, Calif., US), KeyOrganics (Cornwall, UK), Lancaster Synthesis (Windham, N.H., US),Maybridge Chemical Co. Ltd. (Cornwall, UK), Parish Chemical Co. (Orem,Utah, US), Pfaltz & Bauer, Inc. (Waterbury, Conn., US), Polyorganix(Houston, Tex., US), Pierce Chemical Co. (Rockford, Ill., US), Riedel deHaen AG (Hanover, Del.), Spectrum Quality Product, Inc. (New Brunswick,N.J., US), TCI America (Portland, Oreg., US), Trans World Chemicals,Inc. (Rockville, Md., US), and Wako Chemicals USA, Inc. (Richmond, Va.,US).

Methods known to one of ordinary skill in the art are identified throughvarious reference books and databases. Suitable reference books andtreatise that detail the synthesis of reactants useful in thepreparation of compounds described herein, or provide references toarticles that describe the preparation. See, e.g., SYNTHETIC ORGANICCHEM. (John Wiley & Sons, Inc., N.Y.); Sandler et al., ORGANICFUNCTIONAL GROUP PREP., 2nd Ed. (Academic Press, N.Y., 1983); House,MODERN SYNTHETIC REACTIONS, 2nd Ed. (W. A. Benjamin, Inc., Menlo Park,Calif., 1972); Gilchrist, HETEROCYCLIC CHEM., 2nd Ed. (John Wiley &Sons, N.Y., 1992); March, ADVANCED ORGANIC CHEM.: REACTIONS, MECHANISMS& STRUCTURE, 4th Ed., (Wiley-Interscience, N.Y., 1992). Additionalsuitable references that detail the synthesis of reactants useful in thepreparation of compounds described herein, or provide references toarticles that describe the preparation, are known in the art. See, e.g.,Fuhrhop & Penzlin, ORGANIC SYNTH.: CONCEPTS, METHODS, STARTING MAT'LS,2nd Revised & Enlarged Ed. (John Wiley & Sons, ISBN: 3-527-29074-5,1994); HOFFMAN, ORGANIC CHEM., INTERMEDIATE TEXT (Oxford Univ. Press,ISBN 0-19-509618-5, 1996); Larock, COMPREHENSIVE ORGANICTRANSFORMATIONS: GUIDE TO FUNCTIONAL GROUP PREPARATIONS, 2nd Ed.(Wiley-VCH, ISBN: 0-471-19031-4, 1999); March, ADVANCED ORGANIC CHEM.:REACTIONS, MECHANISMS, & STRUCTURE, 4th Ed. (John Wiley & Sons, ISBN:0-471-60180-2, 1992); MODERN CARBONYL CHEM. (Otera (Ed.), Wiley-VCH,ISBN: 3-527-29871-1, 2000); Patai, PATAI'S 1992 GUIDE TO CHEM. OFFUNCTIONAL GROUPS (Interscience ISBN: 0-471-93022-9, 1992); Solomons,ORGANIC CHEM., 7th Ed. (John Wiley & Sons, ISBN: 0-471-19095-0, 2000);Stowell, INTERMEDIATE ORGANIC CHEM., 2nd Ed. (Wiley-Interscience, ISBN:0-471-57456-2, 1993); INDUSTRIAL ORGANIC CHEM.: STARTING MATERIALS &INTERMEDIATES: ULLMANN'S ENCYCLOPEDIA (John Wiley & Sons, ISBN:3-527-29645-X, 1999) in 8 volumes; ORGANIC REACTIONS (1942-2000) (JohnWiley & Sons), in over 55 volumes; CHEM. FUNCTIONAL GROUPS (John Wiley &Sons), in 73 volumes.

Specific and analogous reactants may also be identified through theindices of known chemicals prepared by the Chemical Abstract Service ofthe American Chemical Society, which are available in most public anduniversity libraries, as well as through on-line databases (the AmericanChemical Society, Washington, DC, may be contacted for more details).Chemicals that are known but not commercially available in catalogs maybe prepared by custom chemical synthesis houses, where many of thestandard chemical supply houses (e.g., those listed above) providecustom synthesis services. A reference for the preparation and selectionof pharmaceutical salts of the substituted heterocyclic derivativecompounds described herein is Stahl & Wermuth, HANDBOOK OFPHARMACEUTICAL SALTS (Verlag Helvetica Chimica Acta, Zurich, 2002).

General methods for the synthesis of substituted heterocyclicderivatives are also known. See, e.g., WO 2009158396; WO 200563768; WO2006112666; Briet et. al., 58 Tetrahedron 5761 (2002); WO 200877550; WO200877551; WO 200877556; WO 200712421; WO 200712422; US200799911; WO200877550; Havera et al., 42 J. Med. Chem. 3860 (1999); WO 200429051;US20090054434. Additional examples of the synthesis of substitutedheterocyclic derivatives are known. See, e.g., WO 2012/171337; WO2011/044157; WO 2009/097567; WO 2005/030791; EP 203216; Becknell et al.,21 Bioorg. Med. Chem. Letts. 7076 (2011); Svechkarev et al., VisnikKharkivs'kogo Natsional'nogo Univ. im. V. N. Karazina, 770:201 (2007);Coskun et al., 35 Synth. Commun. 2435 (2005); Alvarez et al., 15 Sci.Synth. 839 (2005); Kihara et al., 53 Heterocycl. 359 (2000); Couture etal., 7 J. Chem. Soc'y, Perkin Transact. 1: Org. Bio-Org. Chem. 789(1999); Kihara et al., 48 Heterocycles 2473 (1998); Couture et al., 52Tetrahed. 4433 (1996); Couturre et al., 37 Tetrahed. Lett. 3697 (1996);Natsugari et al., 38 J. Med. Chem. 3106 (1995); Moehrle et al., 321Archiv Pharm. 759 (Weinheim, Del.) 321:759 (1988); Gore et al., 3 J.Chem. Soc'y, Perkin Transact. 1: Org. Bio-Org. Chem. 481 (1972-1999)(1988); Narasimhan et al., 3 J. Chem. Soc'y, Chem. Commun. 191 (1987);Henry et al., 40 J. Org. Chem. 1760 (1975); Berti, 90 Gazzetta Chim.Italiana 559 (1960); Berti et al., 49 Annal. Chim. 2110 (Rome, Italy)(1959); Berti et al., 49 Annal. Chim. 1253 (Rome, Italy) (1959); WO2012000595; Couture et al., 52 Tetrahed. 4433 (1996); WO 2010069504; WO2010069504; WO 2006030032; WO 2005095384; US20050222159; WO 2013064984;Mishra et al., 2013 Eur. J. Org. Chem. 693 (2013); Vachhani et al., 69Tetrahed. 359 (2013); Xie et al., 45 Eur. J. Med. Chem. 210 (2010);Mukaiyama et al., 15 Bioorg. Med. Chem. 868 (2007); JP2005/089352; Wanget al., 9 Molec. 574 (2004); WO 2000023487; US20060287341; CN103183675;Hares et al., 32 Egyptian J. Pharm. Sci. 303 (1991); DE2356005;DE2133898; DE2133998; U.S. Pat. No. 3,816,422; DE2011970; Staehle etal., 8 Justus Liebigs Annalen der Chem. 1275 (1973).

Additional methods for the synthesis of the substituted heterocyclicderivative compounds disclosed herein are readily available to one ofskill in the art. In some embodiments, the substituted heterocyclicderivative compounds disclosed herein are prepared by the generalsynthetic routes described in the following Schemes 1 and 2, which areexemplary to one of skill in the art and are not limiting.

Referring to Scheme 1, above, 3-fluoro-4-iodopyridine (1-0) undergoes aStille coupling reaction with the tin reagent,tributylstannyl-3,3,3-trifluoro-1-propyne, in presence of a catalyticamount of Pd(0), in toluene, in elevated temperature, such as 130° C.,in a microwave oven to give (1-2). It is then heated with1-azidomethyl-4-methoxy-benzene at reflux in an organic solvent, such ast-butyl alcohol or toluene, to give a mixture ofp-methoxybenzyl-protected (PMB-protected) trifluoromethyl triazoleintermediates (1-4a) and (1-4b). Displacement of 3-fluoro group withamines in presence of K₂CO₃ in dimethyl sulfoxide (DMSO) at hightemperature, such as 180° C., gives mixture (1-5a) and (1-5b), which isthen treated with trifluoroacetic acid (TFA) at room temperature (RT)overnight, or at 50° C. for shorter period of time, to give the finalproduct (1-6).

A method for preparing compounds such as compounds (2-9) is provided inScheme 2, above. According to this approach, 2-fluoro-4-iodopyridine istreated with hydrazine hydrate in an alcoholic solvent, such as ethanol,at elevated temperature (e.g., about 60° C. to 100° C.) to giveintermediate (2-1). Subsequent reaction with an acetoacetyl ester in amixture of an alcoholic solvent (such as ethanol) heated to reflux inpresence of acetic acid provides cyclized hydroxypyrazole pyridineintermediates (2-3). Following a protection of the hydroxyl group byPMB, intermediates (2-4) undergo Stille coupling with the tin reagent,tributylstannyl-3,3,3-trifluoro-1-propyne, in presence of a catalyticamount of Pd(0), in toluene, at elevated temperature, e.g., 120° C., ina microwave oven, to give (2-6). It is then heated with1-azido-methyl-4-methoxy-benzene at reflux in an organic solvent, suchas t-butyl alcohol or toluene, to give a mixture of PMB-protectedtrifluoromethyl triazole intermediates (2-8a) and (2-8b), which aretreated with TFA at room temperature overnight, or at 50° C. for shorterperiod of time, to give the final product (2-9).

In each of the above reaction procedures or schemes, the varioussubstituents may be selected from among the various substituentsotherwise taught herein.

Pharmaceutical Compositions

As noted above, in certain embodiments, the substitutedtriazolylpyridine derivative compounds as described herein may beadministered as a pure chemical or salt thereof. In other embodiments,the substituted triazolylpyridine derivative compounds described hereinare prepared in a pharmaceutical composition in which the substitutedtriazolylpyridine derivative compound is combined with at least onepharmaceutically acceptable or pharmaceutically suitable excipient (alsoreferred to herein as a pharmaceutically suitable (or acceptable)carrier, physiologically suitable (or acceptable) excipient, orphysiologically suitable (or acceptable) carrier), selected on the basisof a chosen route of administration and standard pharmaceuticalpractices, as are well known. See, e.g., REMINGTON: SCI. & PRACTICE PHARM. 21ST ED. (Gennaro, Mack Pub. Co., Easton, Pa., 2005).

Accordingly, provided herein are pharmaceutical compositions thatcomprise at least one substituted heterocyclic derivative compound, or astereoisomer, pharmaceutically acceptable salt, hydrate, solvate, orN-oxide thereof, together with at least one pharmaceutically acceptableexcipient. The excipient (or carrier) is acceptable or suitable if theexcipient is compatible with the other active agents or excipients ofthe composition, not deleterious to the recipient (i.e., the subject) ofthe composition, and prepared under good laboratory practices asrequired for the particular dosage form.

One embodiment provides a pharmaceutical composition comprising acompound of Formula I, or a pharmaceutically acceptable salt thereof.One embodiment provides a pharmaceutical composition comprising acompound of Formula II, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the substituted triazolylpyridine derivativecompounds as described herein are substantially pure, in that suchcompound contains less than about 5%, or less than about 1%, or lessthan about 0.1%, of other organic small molecules, such as contaminatingintermediates or by-products that are created, for example, in one ormore of the steps of a synthesis process.

Suitable oral dosage forms include, for example, tablets, pills,sachets, or capsules of hard or soft gelatin, methylcellulose or ofanother suitable material easily dissolved in the digestive tract.Suitable nontoxic solid carriers are used which include, for example,pharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharin, talcum, cellulose, glucose, sucrose, magnesiumcarbonate, and the like. See, e.g., REMINGTON, 2005.

The dose of the composition comprising at least one substitutedtriazolylpyridine derivative compound as described herein may differ,depending upon the patient's (e.g., human) condition, that is, stage ofthe disease, general health status, age, and other factors that a personskilled in the medical art will use to determine dose.

Pharmaceutical compositions may be administered in a manner appropriateto the disease to be treated (or prevented) as determined by personsskilled in the medical arts. An appropriate dose and a suitable durationand frequency of administration will be determined by such factors asthe condition of the patient, the type and severity of the patient'sdisease, the particular form of the active ingredient, and the method ofadministration. In general, an appropriate dose and treatment regimenprovides the composition(s) in an amount sufficient to providetherapeutic and/or prophylactic benefit (e.g., an improved clinicaloutcome), such as more frequent complete or partial remissions, orlonger disease-free and/or overall survival, or a lessening of symptomseverity. Optimal doses may generally be determined using experimentalmodels and/or clinical trials. The optimal dose may depend upon the bodymass, weight, or blood volume of the patient. Oral doses typically rangefrom about 1.0 mg to about 1000 mg, one to four times, or more, per day.

Histone Demethylase

Chromatin is the complex of DNA and protein that makes up chromosomes.Histones are the major protein component of chromatin, acting as spoolsaround which DNA winds. Changes in chromatin structure are affected bycovalent modifications of histone proteins and by non-histone bindingproteins. Several classes of enzymes are known that can covalentlymodify histones at various sites.

Accordingly, chromatin structure plays a critical role in regulatinggene transcription, which cannot occur efficiently in highly condensedchromatin. Chromatin structure is controlled by a series of posttranslational modifications to histone proteins, notably to histones H3and H4, and most commonly within the “histone tails” which extend beyondthe core nucleosome structure. These post translational modificationsinclude acetylation, methylation, phosphorylation, ribosylationsumoylation, ubiquitination, citrullination, deimination, andbiotinylation. In addition to the histone tails, the cores of histonesH2A and H3 can be modified. Given the function of histones in chromatin,histone modifications are integral to diverse biological processes suchas gene expression, DNA replication, DNA repair, and chromosomecondensation.

Proteins can be post-translationally modified by methylation on aminogroups of lysines and guanidino groups of arginines or carboxymethylatedon aspartate, glutamate, or on the C-terminus of the protein.Post-translational protein methylation has been implicated in a varietyof cellular processes such as RNA processing, receptor mediatedsignaling, and cellular differentiation. Post-translational proteinmethylation is widely known to occur on histones, such reactions knownto be catalyzed by histone methyltransferases, which transfer methylgroups from S-adenyosyl methionine (SAM) to histones. Histonemethylation is known to participate in a diverse range of biologicalprocesses including heterochromatin formation, X-chromosomeinactivation, and transcriptional regulation. Lachner et al., 116 J.Cell Sci. 2117-24 (2003); Margueron et al., 15 Curr. Opin. Genet. Devel.163-76 (2005).

Unlike acetylation, which generally correlates with transcriptionalactivation, whether histone methylation leads to transcriptionactivation or repression depends on the particular site of methylationand the degree of methylation (e.g., whether a particular histone lysineresidue is mono-, di-, or tri-methylated). Methylation of histoneresidues H3K9, H3K27, and H4K20, is generally linked to gene silencing;and methylation of H3K4, H3K36, and H3K79, is generally associated withactive gene expression. In addition, tri- and di-methylation of H3K4generally marks the transcriptional start sites of actively transcribedgenes, whereas mono-methylation of H3K4 is associated withtranscriptional enhancer sequences.

A “demethylase” or “protein demethylase,” refers herein to an enzymethat removes at least one methyl group from an amino acid side chain.Some demethylases act on histones, e.g., act as a histone H3 or H4demethylase. For example, an H3 demethylase may demethylate one or moreof H3K4, H3K9, H3K27, H3K36, or H3K79. Alternately, an H4 demethylasemay demethylate histone H4K20. Demethylases are known that candemethylate either a mono-, di- or a tri-methylated substrate. Further,histone demethylases can act on a methylated core histone substrate, amononucleosome substrate, a dinucleosome substrate, or anoligonucleosome substrate, or a peptide substrate, or chromatin (e.g.,in a cell-based assay).

The first lysine demethylase discovered was lysine-specific demethylase1 (LSD1/KDM1), which uses flavin as a cofactor in demethylating mono-and di-methylated H3K4 or mono- and di-methylated H3K9. A second classof Jumonji C (JmjC) domain-containing histone demethylases werepredicted, and then confirmed when a formaldehyde release assayidentified a H3K36 demethylase; this histone demethylase was named JmjCdomain containing histone demethylase 1 (JHDM1/KDM2A).

Additional JmjC domain-containing proteins were identified subsequently,and they can be phylogenetically clustered into seven subfamilies:JHDM1, JHDM2, JHDM3, JMJD2, JARID, PHF2/PHF8, UTX/UTY, or JmjC domain.

FBXL10 and FBXL11

F-box and leucine-rich repeat protein 10 (FBXL10) and F-box andleucine-rich repeat protein 11 (FBXL11) are multifunctional F-box familyproteins that demethylate histone H3 via a hydroxylation-basedmechanism. FBXL10, also known as lysine (K)-specific demethylase 2B(KDM2B) or Jumonji C domain-containing histone demethylase 1B (JHDM1B),preferentially demethylates trimethylated H3K4 and dimethylated H3K36,but contains weak or no activity for mono- and tri-methylated H3K36.FBXL10 contains three domains: a catalytic JMJC domain, an F-box domain,and a CXXC DNA-binding domain. The N-terminal JMJC domain coordinatesiron and α-ketoglutarate to catalyze demethylation through ahydroxylation-based mechanism. The CXXC DNA-binding domain facilitatesFBXL10 preferential binding to transcribed regions of ribosomal RNA,causing repression of ribosomal RNA gene transcription, which ultimatelycauses inhibition of cell growth and proliferation. FBXL10 isoverexpressed in acute myeloid leukemia, bladder carcinoma, andpancreatic ductal adenocarcinoma. Additionally, FBXL10 regulates theexpression of Polycomb target genes that encode proteins active asepigenetic regulators essential for stem cell differentiation, thusimplicating FBXL10 in tumorigenesis.

FBXL11, also known as KDM2A or JHDM1A, demethylates mono- anddi-methylated H3K36. The FBXL11 CXXC DNA-binding domain recognizesnon-methylated DNA and targets CpG island regions where it specificallyremoves H3K36 methylation. Further, FBXL11 is required to maintain aheterochromatic state, and sustain centromeric integrity and genomicstability during mitosis. In addition, FBXL11 is a key negativeregulator of NF-KB. Overexpression of FBXL11 has been observed innon-small cell lung cancer cell lines (NSCLC), where FBXL11 upregulatesphosphor-ERK1/2 by repressing DUSP3 expression. Negative regulation ofgluconeogenic gene expression by FBXL11 results in suppression of tworate-limiting gluconeogenic enzymes that are critical for maintainingblood glucose homeostasis.

Accordingly, at least one additional embodiment provides a method forinhibiting a histone-demethylase enzyme comprising contacting a histonedemethylase enzyme with a compound of Formula I or Formula II. Anadditional embodiment provides a method of inhibiting ahistone-demethylase enzyme, wherein the histone-demethylase enzymecomprises a JmjC domain. Yet another additional embodiment provides amethod for inhibiting a histone-demethylase enzyme, wherein thehistone-demethylase enzyme is FBXL10 or FBXL11.

Methods of Treatment

Disclosed herein are methods of modulating demethylation in a cell or ina subject, either generally or with respect to one or more specifictarget genes. Demethylation can be modulated to control a variety ofcellular functions, including without limitation: differentiation;proliferation; apoptosis; tumorigenesis, leukemogenesis or otheroncogenic transformation events; hair loss; or sexual differentiation.For example, particular embodiments provide methods of treating adisease regulated by histone methylation or demethylation in a subjectin need thereof by modulating the activity of FBXL10 or FBXL11.

The embodiments further provide a therapeutic method of modulatingprotein methylation, gene expression, cell proliferation, celldifferentiation, or apoptosis in vivo in conditions, illnesses,disorders, infections, or diseases disclosed herein, in particularcancer, inflammatory disease, or viral disease, comprising administeringto a subject in need of such therapy a pharmacologically active ortherapeutically effective amount of at least one substitutedtriazolylpyridine derivative compound described herein, which may beadministered in a pharmaceutical composition.

The embodiments further provide a method of treating a subject, such asa human, suffering from cancer, a neoplastic disease, or otherproliferative disorder. The method comprises administering to a subjectin need of such treatment a therapeutically effective amount of at leastone substituted triazolylpyridine derivative compound described herein,which functions by inhibiting a demethylase and, in general, bymodulating gene expression, to modulate various cellular effects, inparticular inducting or repressing gene expression, arresting cellproliferation, inducing cell differentiation, or inducing apoptosis.

The embodiments further relate to a method for treating or amelioratingcancer, neoplastic disease, or another proliferative disorder mediatedat least in part by histone demethylase activity, by administering aneffective amount of a pharmaceutical composition comprising asubstituted triazolylpyridine derivative compound described herein, to amammal, in particular a human, in need of such treatment. In someaspects, the disease to be treated by the methods of the presentembodiments is cancer.

In a further embodiment is the method for treating cancer in a subjectwherein the cancer pancreatic cancer, prostate cancer, breast cancer,gastric cancer, leukemia, bladder cancer, lung cancer, or melanoma.

Other embodiments and uses will be apparent to one skilled in the art inlight of the present disclosures. The following examples are providedmerely as illustrative of various embodiments and shall not be construedto limit the invention in any way.

EXAMPLES

Unless otherwise noted, reagents and solvents were used as received fromcommercial suppliers. Anhydrous solvents and oven-dried glassware wereused for synthetic transformations sensitive to moisture or oxygen.Yields were not optimized. Reaction times are approximate and were notoptimized. Column chromatography and thin layer chromatography (TLC)were performed on silica gel unless otherwise noted. Spectra are givenin ppm (δ) and coupling constants, J are reported in Hertz. For protonspectra the solvent peak was used as the reference peak.

Preparation 1A: 3-fluoro-4-(3,3,3-trifluoroprop-1-yn-1-yl)pyridine

A mixture of 3-fluoro-4-iodopyridine (1 g, 4.48 mmol),1-tributylstannyl-3,3,3-trifluoro-1-propyne (2.3 g, 5.38 mmol, 90%) andPd(PPh₃)₄ (259 mg, 0.045 mmol) in toluene (15 mL) was heated for 5 hr at130° C. under Na in a microwave oven. The mixture was concentrated andpurified by flash column chromatography on silica gel (EtOAc/Hex=0%-20%)to afford the title compound (260 mg, 31%). [M+H] calculated forC₈H₃F₄N: 190; found: 190.

Preparation 1B:3-fluoro-4-{1-[(4-methoxyphenyl)methyl]-5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl}pyridineand3-fluoro-4-{1-[(4-methoxyphenyl)methyl]-4-(trifluoromethyl)-1H-1,2,3-triazol-5-yl}pyridine

A solution of 3-fluoro-4-(3,3,3-trifluoroprop-1-yn-1-yl)pyridine (260mg, 1.37 mmol) and 1-azidomethyl-4-methoxy-benzene (4 mL, 2 mmol, 0.5Min methyl t-butyl ether) in t-butanol (10 mL) was heated at 80° C. for 2hr. The solution was concentrated and purified by flash columnchromatography on silica gel (EtOAc/Hex=0%-20%) to afford a mixture ofthe title compounds (110 mg, 23%). [M+H] calc'd for C₁₆H₁₂F₄N₄O: 353;found: 353.

Example 1 3-fluoro-4-[5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl]pyridine

A mixture of3-fluoro-4-{1-[(4-methoxyphenyl)methyl]-5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl}pyridineand3-fluoro-4-{1-[(4-methoxyphenyl)methyl]-4-(trifluoromethyl)-1H-1,2,3-triazol-5-yl}pyridine(50 mg, 0.14 mmol) were dissolved in 5 mL of TFA and stirred overnightat 50° C. After the solvent was removed under vacuum, the residue waspurified by flash column chromatography to give the title compound (25mg, 77%). ¹H NMR (400 MHz, DMSO-d6): δ 7.68 (1H, s), 8.62 (s, 1H), 8.81(s, 1H). Calc'd for C₈H₄F₄N₄: 233; found: 233.

Example 2N-(5-chloro-2-fluorophenyl)-4-[5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl]pyridin-3-amine

A mixture of3-fluoro-4-{1-[(4-methoxyphenyl)methyl]-5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl}pyridineand3-fluoro-4-{1-[(4-methoxyphenyl)methyl]-4-(trifluoromethyl)-1H-1,2,3-triazol-5-yl}pyridine(28 mg, 0.08 mmol), 5-chloro-2-fluoroaniline (17 mg, 0.12 mmol) andK₂CO₃ (27 mg, 0.2 mmol) in DMSO (2 mL) was heated at 180° C. for 5 hr ina microwave oven. The reaction mixture was purified by flash columnchromatography on silica gel (EtOAc/Hex=0%-20%) to afford anintermediate which was dissolved in 5 mL of TFA and stirred overnight at50° C. After the solvent was removed under vacuum, the residue waspurified by flash column chromatography to give the title compound (5mg, 17%). ¹H NMR (400 MHz, DMSO-d6): δ 6.81 (1H, dd, J=2.5 and 7.2 Hz),6.90 (1H, m), 7.17 (1H, dd, J=2.6 and 8.7 Hz), 7.40 (1H, d, J=5.0 Hz),8.35 (1H, d, J=5.0 Hz), 8.44 (1H, s). Calculated for C₁₄H₈ClF₄N₅: 359;found: 359.

Example 3N-[(1,2,3,4-tetrahydronaphthalen-1-yl)methyl]-4-[5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl]pyridine-3-amine

The title compound was prepared in 7% yield according to the generalprocedure for the synthesis as in Example 2, starting with Preparation1B. ¹H NMR (400 MHz, DMSO-d6): δ 1.64-1.75 (4H, m), 2.68 (2H, m), 3.08(1H, m), 3.51 (2H, m), 7.09 (3H, m), 7.25 (1H, m), 7.39 (1H, m), 8.01(1H, d, J=4.6 Hz), 8.29 (1H, s). [M+H] calculated for C₁₉H₁₈F₃N₅: 374;found: 374.

Preparation 4A: 2-hydrazinyl-4-iodopyridine

To a solution of 2-fluoro-4-iodopyridine (16 g, 71.74 mmol) in EtOH (160mL) was added NH₂NH₂—H₂O (40 mL). The reaction mixture was stirredovernight at room temp. It was then concentrated in vacuo and theresidue was triturated with PE (200 mL) to afford the desired product(16 g, 95%). [M+H] calculated for C₅H₆IN₃: 235; found: 235.

Preparation 4B:3-[(4-chlorophenyl)methyl]-1-(4-iodopyridin-2-yl)-1H-pyrazol-5-ol

To a solution of ethyl 4-(4-chlorophenyl)-3-oxobutanoate (2 g, 8.30mmol) in EtOH (20 mL) was added a solution of2-hydrazinyl-4-iodopyridine (1.94 g, 8.30 mmol) in EtOH (20 mL) at 50°C., then stirred for 1 hr at 50° C. Then, 5 mL CH₃COOH was added and themixture was refluxed overnight, then concentrated and the residuedissolved in DCM and washed by aq Na₂CO₃, dried, concentrated, andpurified by flash column chromatography on silica gel (PE/EA=5/1 to1/1), to give the title compound (1.9 g, 56%). [M+H] calculated forC₁₅H₁₁ClIN₃O: 412; found: 412.

Preparation 4C:2-{3-[(4-chlorophenyl)methyl]-5-[(4-methoxyphenyl)methoxy]-1H-pyrazol-1-yl}-4-iodopyridine

To a mixture of3-[(4-chlorophenyl)methyl]-1-(4-iodopyridin-2-yl)-1H-pyrazol-5-ol (1 g,2.43 mmol) and K₂CO₃ (470 mg, 3.41 mmol) in DMF (20 mL) was added PMBCl(457 mg, 2.92 mmol) at 0° C., then stirred for 1 hr at 60° C. Afterconcentration, the residue was purified by flash column chromatographyon silica gel (PE/EA=10/1 to 3/1) to give the title compound (430 mg,33%). [M+H] calc'd for C₂₃H₁₉ClIN₃O₂: 532; found: 532.

Preparation 4D:2-{3-[(4-chlorophenyl)methyl]-5-[(4-methoxyphenyl)methoxy]-1H-pyrazol-1-yl}-4-(3,3,3-trifluoroprop-1-yn-1-yl)pyridine

A mixture of2-{3-[(4-chlorophenyl)methyl]-5-[(4-methoxyphenyl)methoxy]-1H-pyrazol-1-yl}-4-iodopyridine(430 mg, 0.81 mmol), 1-tributylstannyl-3,3,3-trifluoro-1-propyne (363mg, 0.85 mmol, 90%) and Pd(Ph₃)₄ (88 mg, 0.081 mmol) in toluene (15 mL)was stirred for 2 hr at 120° C. under Na in a microwave oven. Themixture was concentrated and purified by flash column chromatography onsilica gel (PE/EA=10/1 to 3/1) to afford the title compound (211 mg,52%). [M+H] calculated for C₂₆H₁₉ClF₃N₃O₂: 498; found: 498.

Preparation 4E:2-{3-[(4-chlorophenyl)methyl]-5-[(4-methoxyphenyl)methoxy]-1H-pyrazol-1-yl}-4-{1-[(4-methoxyphenyl)methyl]-5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl}pyridineand2-{3-[(4-chlorophenyl)methyl]-5-[(4-methoxyphenyl)methoxy]-1H-pyrazol-1-yl}-4-{1-[(4-methoxyphenyl)methyl]-4-(trifluoromethyl)-1H-1,2,3-triazol-5-yl}pyridine

A solution of2-{3-[(4-chlorophenyl)methyl]-5-[(4-methoxyphenyl)methoxy]-1H-pyrazol-1-yl}-4-(3,3,3-trifluoroprop-1-yn-1-yl)pyridine(210 mg, 0.42 mmol) and 1-azido-methyl-4-methoxy-benzene (71 mg, 0.42mmol) in toluene (20 mL) was refluxed for 22 hr. The solution wasconcentrated and purified by flash column chromatography on silica gel(PE/EA=10/1 to 3/1) to afford a mixture of the title compounds (226 mg,81%). [M+H] calculated for C₃₄H₂₈ClF₃N₆O₃: 661; found, 661.

Example 43-[(4-chlorophenyl)methyl]-1-{4-[5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl]pyridine-2-yl}-1H-pyrazol-5-ol

A solution of2-{3-[(4-chlorophenyl)methyl]-5-[(4-methoxyphenyl)methoxy]-1H-pyrazol-1-yl}-4-{1-[(4-methoxyphenyl)methyl]-5-(trifluoromethyl)-1H-1,2,3-triazol-4-yl}pyridine(120 mg, 0.18 mmol) in TFA (5 mL) was stirred for 1 hr at 50° C., afterconcentration the residue was purified by prep-HPLC to give the titlecompound (30 mg, 39%). ¹H NMR (400 MHz, DMSO): δ 3.88 (2H, s), 5.24 (1H,br), 7.34-7.51 (6H, m), 8.60 (1H, d, J=5.6 Hz). [M+H] calculated forC₁₈H₁₂ClF₃N₆O: 421; found: 421.

Example 5 In Vitro Enzyme Inhibition Assay

This assay determines the ability of a test compound to inhibit FBXL11,FBXL10, and PHF8 demethylase activity. Baculovirus-expressed FBXL11(GenBank Accession #NM_012308, AA1-1162) was purchased from BPSBioscience (Cat #50102). Baculovirus-expressed FBXL10 (GenBank Accession#NM_032590, AA 1-650) was purchased from BPS Bioscience (Cat #50120).Baculovirus-expressed PHF8 (GenBank Accession NP_055922.1) was purchasedfrom Active Motif (Cat #31435).

FBXL11 Assay

The ability of test compounds to inhibit the activity of FBXL11 wasdetermined in 384-well plate format under the following reactionconditions: 0.15 nM FBXL11, 30 nM H3K36me2-biotin labeled peptide(Anaspec cat #64442), 0.2 μM alpha-ketoglutaric acid in assay buffer of50 mM HEPES, pH7.3, 0.005% Brij35, 0.5 mM TCEP, 0.2 mg/ml BSA, 50 μMsodium L-ascorbate, 5 μM ammonium ironII sulfate. Reaction product wasdetermined quantitatively by AlphaScreen detection after the addition ofdetection reagents anti-H3K36me1 antibody, AlphaScreen®Streptavidin-coated Donor beads, and AlphaScreen® Protein A Acceptorbeads in 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 5 mM EDTA, 2mg/ml BSA to a final 10 μg/ml beads.

The assay reaction was initiated by the following: 3 μl of the mixtureof 90 nM H3K36me2-biotin labeled peptide and 0.6 μM alpha-ketoglutaricacid with 3 μl of 11-point serial diluted inhibitor in 3% DMSO wereadded to each well of 384-well Proxiplate (Perkin Elmer), followed bythe addition of 3 μl of 0.45 nM FBXL11 to initiate the reaction. Thereaction mixture was incubated at room temp for 1 hr, and terminated bythe addition of 3 μl of appropriate dilution of anti-H3K36me1 antibodyin 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 5 mM EDTA, and 2 mg/mlBSA. Plates were then incubated at room temp for 40 min, followed byaddition of 3 μl of 50 μg/ml AlphaScreen® Streptavidin-coated Donorbeads and AlphaScreen® Protein A Acceptor beads in 50 mM HEPES, pH7.3,10 mM NaCl, 0.005% Brij35, 5 mM EDTA, 2 mg/ml BSA. Plates were read byEnVision Multilabel Reader in AlphaScreen mode after minimum 2 hrincubation at room temp. The AlphaScreen signal for each well is used todetermine inhibition constant (IC₅₀).

FBXL10 Assay

The ability of test compounds to inhibit the activity of FBXL10 wasdetermined in 384-well plate format under the following reactionconditions: 0.3 nM FBXL10, 30 nM H3K36me2-biotin labeled peptide(Anaspec cat #64442), 0.2 μM alpha-ketoglutaric acid in assay buffer of50 mM HEPES, pH7.3, 0.005% Brij35, 0.5 mM TCEP, 0.2 mg/ml BSA, 50 μMsodium L-ascorbate, 5 μM ammonium ironII sulfate. Reaction product wasdetermined quantitatively by AlphaScreen detection after the addition ofdetection reagents anti-H3K36me1 antibody, AlphaScreen®Streptavidin-coated Donor beads, and AlphaScreen® Protein A Acceptorbeads in 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 5 mM EDTA, 2mg/ml BSA to a final 10 μg/ml beads.

The assay reaction was initiated by the following: 3 μl of the mixtureof 90 nM H3K36me2-biotin labeled peptide and 0.6 μM alpha-ketoglutaricacid with 3 μl of 11-point serial diluted inhibitor in 3% DMSO wereadded to each well of 384-well Proxiplate (Perkin Elmer), followed bythe addition of 3 μl of 0.9 nM FBXL10 to initiate the reaction. Thereaction mixture was incubated at room temp for 1 hr, and terminated bythe addition of 3 μl of appropriate dilution of anti-H3K36me1 antibodyin 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 5 mM EDTA, and 2 mg/mlBSA. Plates were then incubated at room temp for 40 min, followed byaddition of 3 μl of 50 μg/ml AlphaScreen® Streptavidin-coated Donorbeads and AlphaScreen® Protein A Acceptor beads in 50 mM HEPES, pH7.3,10 mM NaCl, 0.005% Brij35, 5 mM EDTA, 2 mg/ml BSA. Plates were read byEnVision Multilabel Reader in AlphaScreen mode after minimum 2 hrincubation at room temp. The AlphaScreen signal for each well is used todetermine inhibition constant (IC₅₀).

PHF8 Assay

The ability of test compounds to inhibit the activity of PHF8 wasdetermined in 384-well plate format under the following reactionconditions: 3 nM PHF8, 200 nM H3K9me1-biotin labeled peptide (Anaspeccat #64358), 0.5 μM alpha-ketoglutaric acid in assay buffer of 50 mMHEPES, pH7.3, 0.005% Brij35, 0.5 mM TCEP, 0.2 mg/ml BSA, 50 μM sodiumL-ascorbate, and 5 μM ammonium ironII sulfate. Reaction product wasdetermined quantitatively by TR-FRET after the addition of detectionreagent Phycolink Streptavidin-allophycocyanin (Prozyme) andEuropium-anti-unmodified-histone H3 lysine 9/lysine27 (H3K9/K27)antibody (PerkinElmer) in the presence of 5 mM EDTA in LANCE detectionbuffer (PerkinElmer) at a final concentration of 25 nM and 0.5 nM,respectively.

The assay reaction was initiated by the following: 2 μl of the mixtureof 600 nM H3K9me1-biotin labeled peptide and 1.5 μM alpha-ketoglutaricacid with 2 μl of 11-point serial diluted inhibitor in 3% DMSO wereadded to each well of the plate, followed by the addition of 2 μl of 9nM PHF8 to initiate the reaction. The reaction mixture was incubated atroom temp for 15 min, and terminated by the addition of 6 μl of 5 mMEDTA in LANCE detection buffer containing 50 nM PhycolinkStreptavidin-allophycocyanin and 1 nM Europium-anti-unmodified H3K9/K27antibody. Plates were read by EnVision Multilabel Reader in TR-FRET mode(excitation at 320 nm, emission at 615 nm and 665 nm) after 1 hrincubation at RT. A ratio was calculated (665/615) for each well andfitted to determine inhibition constant (IC₅₀).

Example 6 In Vitro Enzyme Inhibition Assay

This assay determines the ability of a test compound to inhibit Jarid1B,JMJD2C and JMJD3 demethylase activity. Baculovirus-expressed Jarid1B(GenBank Accession #NM-006618, AA 2-751) was purchased from BPSBioscience (Cat #50121) or custom made by MolecularThroughput.Baculovirus-expressed JMJD2C (GenBank Accession #BC143571, AA 2-372) waspurchased from BPS Bioscience (Cat #50105). Baculovirus-expressed JMJD3(GenBank Accession #NM-001080424, AA1043-end) was purchased from BPSBioscience (Cat #50115).

Jarid1B Assay

The ability of test compounds to inhibit the activity of Jarid1B wasdetermined in 384-well plate format under the following reactionconditions: 0.8 nM Jarid1B, 300 nM H3K4me3-biotin labeled peptide(Anaspec cat #64357), 2 μM alpha-ketoglutaric acid in assay buffer of 50mM HEPES, pH7.3, 0.005% Brij35, 0.5 mM TCEP, 0.2 mg/ml BSA, 50 μM sodiumL-ascorbate, and 2 μM ammonium ironII sulfate. Reaction product wasdetermined quantitatively by TR-FRET after the addition of detectionreagent Phycolink Streptavidin-allophycocyanin (Prozyme) andEuropium-anti-mono- or di-methylated histone H3 lysine 4 (H3K4me1-2)antibody (PerkinElmer) in the presence of 5 mM EDTA in LANCE detectionbuffer (PerkinElmer) at a final concentration of 25 nM and 1 nM,respectively.

The assay reaction was initiated by the following: 2 μl of the mixtureof 900 nM H3K4me3-biotin labeled peptide and 6 μM alpha-ketoglutaricacid with 2 μl of 11-point serial diluted inhibitor in 3% DMSO was addedto each well of the plate, followed by the addition of 2 μl of 2.4 nMJarid1B to initiate the reaction. The reaction mixture was incubated atroom temp for 30 min, and terminated by the addition of 6 μl of 5 mMEDTA in LANCE detection buffer containing 50 nM PhycolinkStreptavidin-allophycocyanin and 2 nM Europium-anti-H3K4me1-2 antibody.Plates were read by EnVisionMultilabel Reader in TR-FRET mode(excitation at 320 nm, emission at 615 nm and 665 nm) after 1 hrincubation at room temp. A ratio was calculated (665/615) for each welland fitted to determine inhibition constant (IC₅₀).

JMJD2C Assay

The ability of test compounds to inhibit the activity of JMJD2C wasdetermined in 384-well plate format under the following reactionconditions: 0.3 nM JMJD2C, 300 nM H3K9me3-biotin labeled peptide(Anaspec cat #64360), 2 μM alpha-ketoglutaric acid in assay buffer of 50mM HEPES, pH7.3, 0.005% Brij35, 0.5 mM TCEP, 0.2 mg/ml BSA, 50 μM sodiumL-ascorbate, and 2 μM ammonium ironII sulfate. Reaction product wasdetermined quantitatively by TR-FRET after the addition of detectionreagent Phycolink Streptavidin-allophycocyanin (Prozyme) andEuropium-anti-di-methylated histone H3 lysine 9 (H3K9me2) antibody(PerkinElmer) in the presence of 5 mM EDTA in LANCE detection buffer(PerkinElmer) at a final concentration of 50 nM and 1 nM, respectively.

The assay reaction was initiated by the following: 2 μl of the mixtureof 900 nM H3K9me3-biotin labeled peptide and 6 μM alpha-ketoglutaricacid with 2 μl of 11-point serial diluted inhibitor in 3% DMSO wereadded to each well of the plate, followed by the addition of 2 μl of 0.9nM JMJD2C to initiate the reaction. The reaction mixture was incubatedat room temp for 30 min, and terminated by the addition of 6 μl of 5 mMEDTA in LANCE detection buffer containing 100 nM PhycolinkStreptavidin-allophycocyanin and 2 nM Europium-anti-H3K9me2 antibody.Plates were read by EnVisionMultilabel Reader in TR-FRET mode(excitation at 320 nm, emission at 615 nm and 665 nm) after 1 hrincubation at RT. A ratio was calculated (665/615) for each well andfitted to determine inhibition constant (IC₅₀).

MJD3 Assay

The ability of test compounds to inhibit the activity of JMJD3 wasdetermined in 384-well plate format under the following reactionconditions: 1 nM JMJD3, 250 nM H3K27me3-biotin labeled peptide (Anaspeccat #64367), 1 μM alpha-ketoglutaric acid in assay buffer of 50 mMHEPES, pH7.3, 0.005% Brij35, 0.5 mM TCEP, 0.2 mg/ml BSA, 50 μM sodiumL-ascorbate, 5 μM ammonium ironII sulfate. Reaction product isdetermined quantitatively by AlphaScreen detection after the addition ofdetection reagents anti-H3K27me1 antibody, 5 μg/ml AlphaScreen®Streptavidin-coated Donor beads, and 5 μg/ml AlphaScreen® Protein AAcceptor beads in 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 10 mMEDTA, 2 mg/ml BSA.

The assay reaction was initiated by the following: 3 μl of the mixtureof 750 nM H3K27me3-biotin labeled peptide and 3 μM alpha-ketoglutaricacid with 3 μl of 11-point serial diluted inhibitor in 3% DMSO are addedto each well of 384-well Proxiplate (Perkin Elmer), followed by theaddition of 3 μl of 3 nM JMJD3 to initiate the reaction. The reactionmixture was incubated at room temp for 20 min, and terminated by theaddition of 3 μl of appropriate dilution of anti-H3K27me1 antibody in 50mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 10 mM EDTA, 2 mg/ml BSA.Plates were incubated at RT for 1 hr, followed by addition of 3 μl of 25μg/ml AlphaScreen® Streptavidin-coated Donor beads and AlphaScreen®Protein A Acceptor beads in 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005%Brij35, 10 mM EDTA, 2 mg/ml BSA. Plates were read by EnVision MultilabelReader in AlphaScreen mode after minimum 2 hr incubation at room temp.The AlphaScreen signal for each well was used to determine inhibitionconstant (IC₅₀).

The ability to inhibit demethylase activity was quantified and therespective IC₅₀ value was determined for substituted triazolylpyridinederivative compounds. Table 3 provides the IC₅₀ values, in μM, ofvarious compounds disclosed herein; in which the biochemical assay IC₅₀data are designated in the following ranges A: ≦0.10 μM; B: >0.10 μM to≦1.0 μM; C: >1.0 μM to ≦10 μM; and D: >10 μM.

TABLE 3 Chemical Synthesis Example Name FBXL10 JARID1B JMJD2C JMJD3 13-fluoro-4-[5-(trifluoromethyl)-1H-1,2,3- B D D D triazol-4-yl]pyridine2 N-(5-chloro-2-fluorophenyl)-4-[5- C D D D(trifluoromethyl)-1H-1,2,3-triazol-4-yl]pyridin- 3-amine 3N-[(1,2,3,4-tetrahydronaphthalen-1-yl) C D D Dmethyl]-4-[5-(trifluoromethyl)-1H-1,2,3- triazol-4-yl]pyridin-3-amine

Example 7 Preparation of Pharmaceutical Dosage Forms: Oral Tablet

A tablet is prepared by mixing 48% by weight of a compound of Formula Ior Formula II, or a pharmaceutically acceptable salt thereof, 45% byweight of microcrystalline cellulose, 5% by weight of low-substitutedhydroxypropyl cellulose, and 2% by weight of magnesium stearate. Tabletsare prepared by direct compression. The total weight of the compressedtablets is maintained at 250 mg-500 mg.

1. A compound having the structure of Formula I

wherein a compound of Formula I is optionally a pharmaceuticallyacceptable salt thereof, and wherein: R¹ is halogen, CH₂G, NHG, or OG,wherein G is hydrogen, alkyl, or —X—Y, in which X is hydrogen or —C₁alkyl, and Y is optionally substituted aralkyl, aralkenyl, aralkynyl,carbocyclyl, carbocyclylalkyl, heterocyclyl, heterocyclylalkyl,heteroaryl or heteroarylalkyl, or —R^(b)—OR^(a), —R^(b)—OC(O)—R^(a),—R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂, R^(b)—N(R^(a))₂,—R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a), —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a), —R^(b)—S(O)_(t)OR^(a),—R^(b)—S(O)_(t)OR^(a), or —R^(b)—S(O)_(t)N(R^(a))₂, wherein each R^(a)is independently hydrogen, alkyl, fluoroalkyl, cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl, or heteroarylalkyl, each R^(b) is independently a directbond or a straight or branched alkylene or alkenylene chain, each R^(c)is a straight or branched alkylene or alkenylene chain; and t is 1 or 2;and R² is halogen or CF₃; and R⁵ is hydrogen, methyl, ethyl, isoproplyl,t-butyl, CHF₂, CH₂F, CF₃, CH₂OH, CHCH₃OH, or C(CH₃)₂OH.
 2. The compoundof claim 1, wherein R¹ is fluoro.
 3. The compound of claim 1, wherein R²is chloro or fluoro.
 4. The compound of claim 1, wherein R² is CF₃; andR¹ is NHG, wherein G is —X—Y wherein X is methyl.
 5. The compound ofclaim 1, wherein G is —X—Y, and wherein Y is optionally substitutedadamantyl, benzofuranyl, 2,3-dihydrobenzofuranyl, chromanyl, indanyl,indolyl, naphthyl, 1,2-dihydronaphthyl, phenyl, pyridyl,tetrahydroquinolinyl, tetralinyl, 2,3-dihydrobenzo [b][1,4]dioxinyl, orthiochromanyl.
 6. The compound of claim 5, wherein Y is phenylsubstituted with alkyl, alkynyl, chloro, fluoro, fluoroalkyl, nitro, oroptionally substituted aralkyl, aralkenyl, aralkynyl, carbocyclyl,carbocyclylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl orheteroarylalkyl, or —R^(b)—OR^(a), —R^(b)—OC(O)—R^(a),—R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))₂,—R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a), —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a), —R^(b)—S(O)_(t)OR^(a),—R^(b)—S(O)_(t)OR^(a), or —R^(b)—S(O)_(t)N(R^(a))₂; wherein each R^(a)is independently hydrogen, alkyl, fluoroalkyl, cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl, or heteroarylalkyl; each R^(b) is independently a directbond or a straight or branched alkyl or alkenyl chain; each RC is astraight or branched alkyl or alkenyl chain; and t is 1 or
 2. 7. Thecompound of claim 1, wherein the compound has the structure:

wherein R², R⁵, and G are as described in claim
 1. 8. The compound ofclaim 1, wherein the compound has the structure:

wherein R², and R⁵ are as described in claim 1; and Z is independentlyat least one hydrogen, hydroxy, halogen, cyano, NH₂, or optionallysubstituted alkyl, alkenyl, alkynyl, alkoxy, aryl, aryl-S—, aralkyl,aryloxy, aralkoxy, carbocyclyl, carbocyclylalkyl, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclylalkyl or heteroarylalkyl, orNHR^(d), N(R^(d))₂, NHC(O)R^(d), NHC(O)OR^(d), NHC(O)NHR^(d),NHC(O)N(R^(d))₂, NHS(O)₂R^(d), NR^(d)C(O)R^(d), NR^(d)C(O)OR^(d),NR^(d)C(O)NHR^(d), NR^(d)C(O)N(R^(d))₂, or NR^(d)S(O)₂R^(d), whereineach R^(d) is independently alkyl, aryl, aralkyl, carbocyclyl,heterocyclyl, heteroaryl, carbocyclylalkyl, heterocyclylalkyl, orheteroarylalkyl.
 9. The compound of claim 8, wherein R² is CF₃; and Z isindependently at least one fluoro, phenylmethoxy, trifluoromethyloxy, oroptionally substituted straight, branched or cyclic C₁-C₆ alkyl.
 10. Thecompound of claim 8, wherein R² is CF₃, one of Z is fluoro, and one of Zis NHR^(d) or N(R^(d))₂.
 11. The compound of claim 1, having thestructure:


12. A compound having the structure of Formula II

wherein a compound of Formula II is optionally a pharmaceuticallyacceptable salt thereof, and wherein R² is halogen or CF₃; R³ ishydrogen, halogen, —OH, —OR⁶, —N(R⁶)₂, or optionally substituted alkyl,carbocyclyl, heterocyclyl, aryl, heteroaryl, carbocyclylalkyl,heterocyclylalkyl, aralkyl, or heteroarylalkyl; R⁴ is hydrogen, halogen,—OH, —OR⁶, —N(R⁶)₂, or optionally substituted alkyl, carbocyclyl,heterocyclyl, aryl, heteroaryl, carbocyclylalkyl, heterocyclylalkyl,aralkyl, or heteroarylalkyl; wherein each R⁶ is independently hydrogen,or optionally substituted alkyl, carbocyclyl, heterocyclyl, aryl,heteroaryl, carbocyclylalkyl, heterocyclylalkyl, aralkyl, orheteroarylalkyl; and R⁵ is hydrogen, methyl, ethyl, isoproplyl, t-butyl,CHF₂, CH₂F, CF₃, CH₂OH, CHCH₃OH, or C(CH₃)₂OH.
 13. The compound of claim12, wherein R² is chloro or fluoro.
 14. The compound of claim 12,wherein R⁴ and R⁵ are hydrogen, and R³ is optionally substitutedaralkyl, aryloxyalkyl, cyclylalkyl, or heterocyclylalkyl.
 15. Thecompound of claim 12, having the structure:


16. A method of inhibiting a histone demethylase enzyme comprisingcontacting the histone demethylase enzyme with the compound of claim 1.17. A pharmaceutical composition comprising the compound of claim
 1. 18.A method of treating cancer or other disease associated with abnormalhistone demethylase activity in a subject in need of such treatment,comprising administering to the subject the pharmaceutical compositionof claim
 17. 19. A method of inhibiting a histone demethylase enzymecomprising contacting the histone demethylase enzyme with the compoundof claim
 12. 20. A pharmaceutical composition comprising the compound ofclaim
 12. 21. A method of treating cancer or other disease associatedwith abnormal histone demethylase activity in a subject in need of suchtreatment, comprising administering to the subject the pharmaceuticalcomposition of claim 20.